RU2806320C2 - Aqueous pharmaceutical composition of antibody to pd-1 of prolgolimab and its use - Google Patents

Abstract

FIELD: pharmaceuticals.

SUBSTANCE: group of inventions relates to aqueous pharmaceutical compositions for anti-PD-1 antibody prolgolimab, to the use of such pharmaceutical compositions and to a method of treatment using them. Variants of an aqueous pharmaceutical composition of anti-PD-1 antibodies are proposed, consisting of the following: prolgolimab as an antibody, trehalose dihydrate, sodium acetate trihydrate in certain amounts and acetic acid to provide pH in the range of 5.0–5.5; as well as variants of the aqueous pharmaceutical composition of anti-PD-1 antibodies, consisting of the following: prolgolimab as an antibody, trehalose dihydrate, L-histidine and L-histidine hydrochloride in certain amounts, at pH in the range of 5.5–6.0. The qualitative and quantitative composition of the excipients used in the composition variants ensures the stability of prolgolimab aqueous compositions in a wide range of its concentrations. Also provided is the use of the above variants of the aqueous pharmaceutical composition of the anti-PD-1 antibody of prolgolimab for the treatment of a malignant neoplasm, and a method for the treatment of a malignant neoplasm comprising administering a therapeutically effective amount of the variants of the above aqueous pharmaceutical composition.

EFFECT: obtaining aqueous pharmaceutical composition of antibody to PD-1 of prolgolimab.

58 cl, 23 tbl, 19 ex, 12 dwg

Description

Field of technology

[001] The present invention relates to new aqueous compositions for anti-PD-1 antibodies, and, in particular, to new aqueous compositions for the anti-PD-1 antibody prolgolimab, which can be used as a drug for the treatment of cancer.

State of the art

[002] Programmed death protein 1 (PD-1) is an inhibitory member of the CD28 receptor family, which also includes CD28, CTLA-4, ICOS and BTLA. PD-1 is expressed by activated B cells, T cells, and myeloid cells (Agata et al., supra; Okazaki et al. (2002) Curr. Opin. Immunol. 14: 391779-82; Bennet et al. (2003) J Immunol 170:711-8). The original members of this family, CD28 and ICOS, were found to function in increasing T cell proliferation following the addition of monoclonal antibodies (Hutloff et al. (1999) Nature 397:263-266; Hansen et al. (1980) Immunogenics 10:247 -260). PD-1 was discovered by screening for differential expression in apoptotic cells (Ishida et al. (1992) EMBO J 11:3887-95). Other members of this family, CTLA-4 and BTLA, were detected by screening for differential expression in cytotoxic T lymphocytes and TH1 cells, respectively. CD28, ICOS, and CTLA-4 all have an unpaired cysteine residue allowing homodimerization. In contrast, PD-1 is predicted to exist as a monomer, lacking the unpaired cysteine residue characteristic of other CD28 family members.

[003] PD-1 is a 55 kDa type I transmembrane protein that is part of the Ig gene superfamily (Agata et al. (1996) Int Immunol 8:765-72). PD-1 contains a membrane-proximal immunoreceptor tyrosine-based inhibitory motif (ITIM) and a membrane-distal tyrosine-based switch motif (ITSM) (Thomas, M.L. (1995) J Exp Med 181:1953-6; Vivier, E and Daeron, M (1997) Immunol Today 18:286-91). PD-1, although structurally similar to CTLA-4, lacks the MYPPPY motif, which is critical for the binding of B7-1 and B7-2. Two ligands for PD-1, PD-L1 and PD-L2, have been identified and have been shown to negatively regulate T cell activation upon binding to PD-1 (Freeman et al. (2000) J Exp Med 192:1027- 34;Latchman et al (2001) Nat Immunol 2:261-8;Carter et al (2002) Eur J Immunol 32:634-43). Both PD-L1 and PD-L2 are B7 homologs that bind to PD-1 but not to other CD28 family members.

[004] One ligand for PD-1, PD-L1, is abundant in various types of human cancer (Dong et al. (2002) Nat. Med. 8:787-9). The interaction between PD-1 and PD-L1 leads to a decrease in the number of tumor-infiltrating lymphocytes, a decrease in T-cell receptor-mediated proliferation, and immune evasion of cancer cells (Dong et al. (2003) J. Mol. Med. 81:281-7 ; Blank et al. (2005) Cancer Immunol. Immunother. 54:307-314; Konishi et al. (2004) Clin. Cancer Res. 10:5094-100). Immunosuppression can be reversed by inhibiting the local interaction of PD-1 with PD-L1, and this effect is additive when blocking the interaction of PD-1 with PD-L2 (Iwai et al. (2002) Proc. Nat'l. Acad. Sci. USA 99 :12293-7; Brown et al (2003) J Immunol 170:1257-66).

[005] PD-1 is an inhibitory member of the CD28 family expressed on activated B cells, T cells and myeloid cells (Agata et al., supra; Okazaki et al. (2002) Curr Opin Immunol 14: 391779-82; Bennett et al (2003) J Immunol 170:711-8). PD-1-deficient animals develop various autoimmune phenotypes, including autoimmune cardiopathy and a lupus-like syndrome with arthritis and nephritis (Nishimura et al. (1999) Immunity 11:141-51; Nishimura et al. (2001) Science 291:319-22 ). Additionally, PD-1 has been found to play a role in autoimmune encephalomyelitis, systemic lupus erythematosus, graft-versus-host disease (GVHD), type I diabetes, and rheumatoid arthritis (Salama et al. (2003) J Exp Med 198:71-78 ; Prokunina and Alarcon-Riquelme (2004) Hum Mol. Genet 13:R143; Nielsen et al. (2004) Lupus 13:510). In the murine tumor B cell line ITSM, PD-1 has been shown to be essential for blocking BCR-mediated Ca2+ influx and tyrosine phosphorylation of downstream effector molecules (Okazaki et al. (2001) PNAS 98:13866-71) .

[006] Currently, a number of antibodies against PD-1 are known, for example, nivolumab (BMS), pembrolizumab (Merck), which are a human monoconal antibody of the IgG4 isotype.

[007] Also known is a new anti-PD-1 antibody, prolgolimab (also known as BCD-100), which is a human monoclonal antibody of the IgG1 isotype with effectorless mutations L234A, L235A. Prolgolimab showed increased affinity for PD-1 and increased aggregation stability compared to antibodies of the IgG4 isotype. In addition, prolgolimab is currently in clinical trials for various types of malignancies, including melanoma, including unresectable or metastatic melanoma, early stage melanoma before and after curative treatment; lung cancer, non-small cell lung cancer (NSCLC), including inoperable or metastatic non-small cell lung cancer.

[008] Thus, the development of new and improved stable aqueous pharmaceutical compositions for the anti-PD-1 antibody prolgolimab is currently relevant.

Brief description of drawings

[009] The invention will become better understood from the following detailed description of embodiments of the present invention with reference to the accompanying drawings, in which:

[0010] Figure 1, Figure 2 are graphs illustrating the dynamics of BCD-100 concentrations in the blood serum of patients over 6 administrations of the drug (taking into account the coefficient) in μg/ml. (Study BCD-100-1).

[0011] Figure 3 is a graph illustrating the design of the BCD-100-2/MIRACULUM study.

[0012] Figure 4 is a graph illustrating a flowchart of the study.

[0013] Figure 5 is a graph illustrating the overall survival of patients in group 1 (BCD-100, 1 mg/kg every 2 weeks) from the BCD-100-2/MIRACULUM study.

[0014] Figure 6 is a graph illustrating the overall survival of patients in group 2 (BCD-100, 3 mg/kg every 3 weeks) from the BCD-100-2/MIRACULUM study.

[0015] Figure 7 is a graph illustrating the progression-free survival of patients in the BCD-100, 1 mg/kg group (according to irRECIST criteria) from the BCD-100-2/MIRACULUM study.

[0016] Figure 8 is a graph illustrating the progression-free survival of patients in the BCD-100 3 mg/kg group (according to irRECIST criteria) from the BCD-100-2/MIRACULUM study.

[0017] Figure 9 is a graph illustrating the concentration of BCD-100 in patients receiving 1 mg/kg every 2 weeks after administration of a single dose of the drug (results of the BCD-100-2 /MIRACULUM study).

[0018] Figure 10 is a graph illustrating the concentration of BCD-100 in patients receiving 3 mg/kg every 3 weeks (results from the BCD-100-2/MIRACULUM study).

[0019] Figure 11 is a graph illustrating the proportion of Th9 in the total T helper cell population in subgroups of patients treated with BCD-100 at a dose of 1 mg/kg Q2W, with different types of response to therapy (results of the BCD-100-2 /MIRACULUM study ).

[0020] Figure 12 is a graph illustrating the proportion of Th9 in the total T helper cell population in subgroups of patients treated with BCD-100 at a dose of 3 mg/kg Q3W, with different types of response to therapy (results of the BCD-100-2 /MIRACULUM study ).

Description of the invention Definitions:

[0021] The terms used herein generally have their common art-specific meanings within the context of the invention and in the particular context in which each term is used. Below, or elsewhere in the description, certain terms that are used to describe the invention are defined to provide the practitioner with additional guidance regarding the description of the invention. Synonyms of some terms are given. The presentation of one or more synonyms does not exclude the use of other synonyms. The use of examples anywhere herein, including examples of any terms described herein, is for illustrative purposes only and does not in any way limit the scope or meaning of the invention or any subject matter illustrated. The invention is not limited to the various embodiments described in the present description.

[0022] "Monoclonal antibody" as used herein refers to a humanized antibody or a fully human antibody unless otherwise specified in this application. Monoclonal antibodies of the invention can be produced using, for example, recombinant technologies, phage display technologies, synthetic technologies or combinations of such technologies or other technologies well known in the art.

[0023] A "monoclonal antibody" population, as used herein, refers to a homogeneous or substantially homogeneous population of antibodies (i.e., at least about 96%, but more preferably at least about 97 or 98% or more preferably at least 99% of the antibodies in the population will compete in the ELISA for the same antigen or epitope, or more preferably the antibodies are identical in amino acid sequence).

[0024] A full-length antibody that exists in nature is an immunoglobulin molecule that consists of four polypeptide chains (two heavy (H) chains (approximately 50-70 kDa at full length) and two light (L) chains (approximately 25 kDa at full length) full length)), connected by disulfide bridges. The amino-terminal portion of each chain includes a variable domain of approximately 100-110 or more amino acids that is responsible for antigen binding. The carboxy-terminal portion of each chain defines a constant region primarily responsible for effector function. Light chains are classified as kappa or lambda and are characterized by a specific constant region. Each light chain consists of an N-terminal light chain variable region (in this application "VL" or "VK") and a light chain constant region consisting of a single domain (CL or CK). Heavy chains are classified as γ (gamma), δ (delta), alpha (α), mu (μ) or epsilon (ε), and they define the isotype of the antibody, such as IgG, IgM, IgA, IgD and IgE, respectively, and several of them can be further divided into subclasses (isotypes) such as IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2. Each type of heavy chain is characterized by a specific Fc constant region. Each heavy chain consists of an N-terminal heavy chain variable region (in this application "VH") and a constant region CH (heavy chain). The heavy chain constant region consists of three domains (CH1, CH2 and CH3) for IgG, IgD and IgA and 4 domains (CH1, CH2, CH3 and CH4) for IgM and IgE. The VH and VL variable domains can be further divided into hypervariable regions (hypervariable regions, CDRs) interspersed with more conserved framework regions (FRs). Each variable domain consists of three CDRs and four FRs, arranged in the following order from N-terminus to C-terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3 and FR4.

[0025] The variable regions of each of the light/heavy chain pairs form the antigen-binding sites of the antibody. Thus, an intact IgG antibody has two binding sites. With the exception of bifunctional or bispecific antibodies, the two binding sites are the same. As used herein, an “antigen-binding portion” or “antigen-binding region” or “antigen-binding domain” refers, interchangeably, to that part of an antibody molecule that contains amino acid residues that interact with the antigen and determine the specificity and affinity of the antibody for the antigen. . This part of the antibody includes the "framework" amino acid residues necessary to maintain the proper conformation of the antigen-binding residues.

[0026] An "antibody fragment" may be an antibody fragment or an antibody fragment having the activity of a full-length antibody. Said antibody fragment may be F(ab')2, F(ab)2, Fab', Fab Fv and scFv.

[0027] The term “inhibit” or “neutralize,” as used herein, with respect to the functional activity of an antibody of the invention, means the ability to substantially inhibit, prevent, limit, delay, terminate, reduce or reverse, for example, the development or the severity of what is inhibited, including, but not limited to, the biological activity (eg, PD-1 activity) or property, disease or condition. The inhibition or neutralization of PD-1 activity resulting from binding of an antibody of the invention to PD-1 is preferably at least about 20, 30, 40, 50, 60, 70, 80, 90, 95% or greater.

[0028] The term "isolated" or "isolated" when used with respect to a nucleic acid or protein preparation (eg, an antibody) refers to a nucleic acid molecule or protein molecule that is identified and separated from at least one contaminant with which it usually bound in a natural source. Preferably, the "isolated antibody" is an antibody that is substantially free of other antibodies having distinctive antigen specificity (e.g., the pharmaceutical compositions of the present invention contain an isolated antibody that specifically binds PD-1 and is substantially free of antibodies that specifically bind antigens other than PD-1).

[0029] A polynucleotide is “operably linked” if it is functionally linked to another polynucleotide. For example, a promoter or enhancer is operably linked to a coding sequence if it affects transcription of the sequence. A polypeptide is "operably linked" to another polypeptide if the polynucleotides encoding them are operably linked, preferably if they are in the same open reading frame.

[0030] The term "Specific binding" between an antibody and an antigen target (antigen) refers to immunological specificity. An antibody can specifically bind to a target antigen if it binds an epitope on the antigen to a greater extent than other epitopes on the antigen. Specific binding does not exclude cross-reactivity with other antigens bearing similar antigen epitopes.

[0031] VL domains in antibodies according to the invention may be of the VL lambda type, or of the VL kappa type. The term "VL domain" refers to both VK lambda and VL kappa isotypes that contain one or more amino acid substitutions, insertions or deletions.

[0032] The term "pharmaceutical composition" refers to a composition and/or composition containing an antibody according to the invention in a therapeutically effective amount and excipients or excipients (carriers, diluents, fillers, solvents and other excipients).

[0033] As used herein, the term "buffer" or "buffer solution" refers to an aqueous solution containing a mixture of an acid (usually a weak acid, such as, for example, acetic acid, citric acid) and its conjugate base (such as, for example, acetate or citrate salt, for example sodium acetate, sodium citrate, and hydrates of these salts, for example sodium acetate trihydrate) or alternatively a mixture of a base (usually a weak base, for example histidine) and its conjugate acid (for example histidine hydrochloride). The pH value of a "buffer solution" changes little when a small amount of a strong base or a strong acid is added to it, or when diluted or concentrated, due to the "buffering effect" provided by the "buffering agent".

[0034] As used herein, a "buffer system" contains one or more buffering agents and/or their acid or base conjugate(s), and more suitably contains one or more buffering agents and their acid or base conjugate(s), and most suitably contains only one buffering agent and its acid/base conjugate. Unless otherwise indicated, any concentrations referred to in the present invention with respect to a "buffer system" (buffer concentration) may refer to the combined concentration of the buffer(s) agent(s) and/or its acid or base conjugate(s). In other words, the concentrations referred to herein with respect to a “buffer system” may refer to the combined concentration of the relevant buffer species (ie, species in dynamic equilibrium with each other, eg, citrate/citric acid). The total pH of a composition containing a relevant buffer system is a reflection of the equilibrium concentration of each of the relevant buffer species (ie, the balance of the buffer(s) agent(s) with its acid or base conjugate(s).

[0035] As used herein, the term "buffering agent" refers to the acidic or alkaline component (usually a weak acid or weak base) of a buffer or buffer solution. A buffering agent helps maintain the pH of a given solution at or near a predetermined value, and buffering agents are typically selected to complement the predetermined value. The buffering agent may be a single compound that produces the desired buffering effect, particularly if said buffering agent is mixed with (and suitably proton exchangeable with) a suitable amount (depending on the predetermined desired value) of its corresponding "acid/base" conjugate".

[0036] The term "solubilizer" as used herein means a pharmaceutically acceptable non-ionic surfactant. A single solubilizer as well as combinations of solubilizers can be used. Examples of solubilizers include, but are not limited to, polysorbate 20 or polysorbate 80, poloxamer 184 or poloxamer 188, or ^PLURONIC® .

[0037] The terms "osmotic agent" or "tonicity-regulating agent" and osmolytic as used herein refer to an excipient that can provide the desired osmotic pressure to a liquid antibody solution. In some embodiments, the tonicity adjusting agent can adjust the osmotic pressure of the liquid antibody preparation to isotonicity such that the antibody preparation is physiologically compatible with the tissue cells of the subject. In yet another embodiment, the "tonicity-regulating agent" may help increase the stability of the antibodies. An "isotonic" drug is a drug that has an osmotic pressure equivalent to human blood. Isotonic drugs typically have an osmotic pressure of approximately 250 to 350 mOsm/kg. The term "hypotonic" describes a drug with an osmotic pressure less than that of human blood. Accordingly, the term "hypertonic" is used to describe a drug with an osmotic pressure greater than that of human blood. Isotonicity can be measured using, for example, a steam or cryoscopic osmometer. The tonicity adjusting agent may be in enantiomeric (eg, L- or D-enantiomer) or racemic form; in the form of isomers such as alpha or beta, including alpha, alpha; or beta, beta; or alpha, beta; or beta, alpha; in free acid or free base form; in the form of salt; in hydrated form (eg monohydrate) or anhydrous form. Examples of osmotic agents include, but are not limited to, sugars (trehalose dihydrate, sucrose, glucose), polyols (mannitol, sorbitol), amino acids (proline, arginine, glycine), or salts (sodium chloride, potassium chloride, magnesium chloride).

[0038] The term "long-term storage" or "long-term stability" should be understood to mean that the pharmaceutical composition can be stored for three months or more, for six months or more, and preferably for one year or more, most preferably with a minimum shelf life in stable condition of at least two years. In general, the terms “long-term storage” and “long-term stability” further include steady state storage times that are at least comparable to or better than the steady state storage times typically required for currently available commercial anti-PD antibody formulations -1 prolgolimab without any loss in stability that would render the formulation unsuitable for its intended pharmaceutical use.

[0039] The term "parenteral administration" means modes of administration, typically by injection, and includes, but is not limited to, intravenous, intramuscular, intraarterial, intratracheal, intracapsular, intraorbital, intracardial, intradermal, intraperitoneal, transtracheal, subcutaneous, intraarticular, subcapsular, subarachnoid , intraspinal, epidural and epigastric injection or infusion.

[0040] The term "use" refers to the possibility of using an antibody according to the invention or a pharmaceutical composition containing it, for the treatment, alleviation of diseases, to accelerate remission, reduce the frequency of relapses due to diseases or disorders mediated by receptors to which the antibody according to the invention can bind . Examples of diseases include, but are not limited to, malignancies, including melanoma, including unresectable or metastatic melanoma, early stages of melanoma before and after radical treatment; lung cancer, non-small cell lung cancer (NSCLC), including unresectable or metastatic non-small cell lung cancer; non-squamous non-small cell lung cancer, squamous cell lung cancer; small cell lung cancer, including unresectable or metastatic small cell lung cancer; early stage lung cancer before and after radical treatment; cervical cancer, including metastatic cervical cancer, early stages of cervical cancer before and after radical treatment; head and neck tumors, including squamous cell carcinoma of the head and neck; Hodgkin's lymphoma; tumors of the stomach and intestines, metastatic squamous cell carcinoma of the esophagus; bladder cancer, including metastatic urothelial carcinoma, kidney cancer; endometrial cancer, including metastatic endometrial cancer, early stages of endometrial cancer before and after radical treatment; breast cancer, including metastatic breast cancer, early stages of endometrial cancer before and after radical treatment; liver cancer, including metastatic or inoperable liver cancer, early stages of liver cancer before and after radical treatment; unresectable or metastatic solid tumor, including unresectable or metastatic solid tumor with signs of microsatellite instability.

[0041] The term “method of treatment” refers to the possibility of using an antibody according to the invention or a pharmaceutical composition containing it, for the treatment, alleviation of diseases, to accelerate remission, reduce the frequency of relapses due to diseases or disorders associated with PD1 activity. “Treat” or “treating” a disease, disorder or condition may include preventing or delaying the onset of clinical symptoms of the disease, disorder or condition developing in a person, inhibiting the disease, disorder or condition, that is, stopping, reducing or slowing the progression of the disease or its relapse (in the case of maintenance therapy) or at least one clinical or subclinical symptom thereof, or alleviation or attenuation of the disease, that is, causing regression of the disease, disorder or condition. Examples of diseases include, but are not limited to, malignancies, including melanoma, including unresectable or metastatic melanoma, early stages of melanoma before and after radical surgical treatment; lung cancer, non-small cell lung cancer (NSCLC), including unresectable or metastatic non-small cell lung cancer.

[0042] The term "aqueous composition" as used herein refers to a water-based composition, the water may include: water, water for injection, saline solution (0.9-1.0% aqueous sodium chloride solution ).

[0043] In one embodiment of the invention, the subject of treatment or patient is a mammal, preferably a human subject. The above subject may be male or female of any age.

[0044] In the present specification and in the following claims, unless the context otherwise requires, the words “have”, “include” and “comprise” or variations thereof such as “has”, “having”, “includes”, “comprising” ", "contains" or "containing" shall be understood to mean the inclusion of the specified whole or group of wholes, but not the exclusion of any other whole or group of wholes.

The essence of the invention

[0045] The present invention discloses aqueous pharmaceutical compositions of the anti-PD-1 antibody prolgolimab, which has improved aggregation stability, increased affinity compared to known anti-PD-1 antibodies that are based on the human IgG4 isotype antibody.

[0046] As described in international patent application publication WO/2018/013017, incorporated herein by reference, the anti-PD-1 antibody prolgolimab, which is a human monoclonal antibody of the IgG1 isotype with the effectorless mutations L234A, L235A, (referred to herein as the “antibody of the invention”) has improved aggregation stability, increased affinity, and improved pharmacokinetic parameters such as t1/2β (hour) or Cmax (μg/ml) compared to known anti-PD-1 antibodies, which are based on human antibodies of the IgG4 isotype, such as nivolumab. Prolgolimab has a weight average molecular mass of approximately 14 6 kDa and is specific for human PD-1. Prolgolimab has a heavy chain of 459 amino acids (SEQ ID NO: 1) and a human lung of 214 amino acids (SEQ ID NO: 2), the constant portion (Fc) of prolgolimab contains mutations L234A, L235A.

[0047] In this regard, it would be useful to administer an aqueous antibody composition of the invention to a patient with cancer.

[0048] A broad aspect of the present invention is an aqueous pharmaceutical composition suitable for administration to a subject to inhibit the activity of the PD-1 protein. The aqueous pharmaceutical composition contains a pharmaceutically effective amount of the anti-PD-1 antibody prolgolimab, an effective amount of trehalose dihydrate, a buffering agent based on acetate or histidine.

[0049] In accordance with one broad aspect of the present invention, there is provided an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

(a) prolgolimab at a concentration of 15 mg/ml to 40 mg/ml as an antibody;

(b) trehalose dihydrate at a concentration of 80 mg/ml to 110 mg/ml;

(c) sodium acetate trihydrate at a concentration of 0.2 mg/ml to 2.5 mg/ml; And

(d) acetic acid to pH 4.5 to 5.5.

[0050] In some embodiments, said prolgolimab may be at a concentration of 15 mg/ml to 25 mg/ml.

[0051] In some embodiments, said prolgolimab may be at a concentration of 20 mg/ml.

[0052] In some embodiments, said trehalose dihydrate may be present at a concentration of from 95 mg/ml to 105 mg/ml.

[0053] In some embodiments, said trehalose dihydrate may be present at a concentration of 100 mg/ml.

[0054] In some embodiments, said sodium acetate trihydrate may be present at a concentration of 1.6 mg/ml to 1.9 mg/ml.

[0055] In some embodiments, said sodium acetate trihydrate may be present at a concentration of 1.7 mg/ml to 1.8 mg/ml.

[0056] In some embodiments, said sodium acetate trihydrate may be present at a concentration of 1.742 mg/ml.

[0057] In some embodiments, said acetic acid may be added to pH 5.0.

[0058] In some embodiments, said acetic acid may be present at a concentration of 0.04 to 0.77 mg/ml.

[0059] In some embodiments, said acetic acid may be present at a concentration of 0.40 mg/ml to 0.50 mg/ml.

[0060] In some embodiments, said acetic acid may be present at a concentration of 0.43 mg/ml.

[0061] In accordance with one broad aspect of the present invention, there is provided an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

(a) prolgolimab at a concentration of 90 mg/ml to 150 mg/ml as an antibody;

(b) trehalose dihydrate at a concentration of 50 mg/ml to 110 mg/ml;

(c) sodium acetate trihydrate at a concentration of 0.2 mg/ml to 2.5 mg/ml; And

(d) acetic acid to pH 4.5 to 5.5.

[0062] In some embodiments, said prolgolimab may be at a concentration of 90 mg/ml to 110 mg/ml.

[0063] In some embodiments, said prolgolimab may be present at a concentration of 100 mg/ml.

[0064] In some embodiments, said trehalose dihydrate may be present at a concentration of 75 mg/ml to 85 mg/ml.

[0065] In some embodiments, said trehalose dihydrate may be present at a concentration of 80 mg/ml.

[0066] In some embodiments, said sodium acetate trihydrate may be present at a concentration of from 1.6 mg/ml to 1.9 mg/ml.

[0067] In some embodiments, said sodium acetate trihydrate may be present at a concentration of 1.7 mg/ml to 1.8 mg/ml.

[0068] In some embodiments, said sodium acetate trihydrate may be present at a concentration of 1.742 mg/ml.

[0069] In some embodiments, said acetic acid may be added to a pH of 5.0 to 5.5.

[0070] In some embodiments, said acetic acid may be added to pH 5.0.

[0071] In some embodiments, said acetic acid may be present at a concentration of 0.045 mg/ml to 0.77 mg/ml.

[0072] In some embodiments, said acetic acid may be present at a concentration of 0.40 mg/ml to 0.50 mg/ml.

[0073] In some embodiments, said acetic acid may be present at a concentration of from 0.4 to 3 mg/ml.

[0074] In accordance with one broad aspect of the present invention, there is provided an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

(a) prolgolimab at a concentration of 5 mg/ml to 150 mg/ml as an antibody;

(b) trehalose dihydrate at a concentration of 70 mg/ml to 110 mg/ml;

(c) L-histidine at a concentration of 0.2 to 2.5 mg/ml; And

(d) L-histidine hydrochloride at a concentration of 0.2 to 3.5 mg/ml.

[0075] In some embodiments, said prolgolimab may be at a concentration of 15 mg/ml to 40 mg/ml.

[0076] In some embodiments, said prolgolimab may be at a concentration of 15 mg/ml to 25 mg/ml.

[0077] In some embodiments, said prolgolimab may be present at a concentration of 20 mg/ml.

[0078] In some embodiments, said trehalose dihydrate may be present at a concentration of from 95 mg/ml to 105 mg/ml.

[0079] In some embodiments, said trehalose dihydrate may be present at a concentration of 100 mg/ml.

[0080] In some embodiments, said L-histidine may be present at a concentration of 0.7 to 1.0 mg/ml.

[0081] In some embodiments, said L-histidine may be present at a concentration of 0.92 mg/ml.

[0082] In some embodiments, said L-histidine hydrochloride may be present at a concentration of 2.8 to 3.3 mg/ml.

[0083] In some embodiments, said L-histidine hydrochloride may be present at a concentration of 2.96 mg/ml.

[0084] In some embodiments, the composition may have a pH of 5.5 to 6.5.

[0085] In some embodiments, the composition may have a pH of 5.5.

[0086] In some embodiments, said prolgolimab may be at a concentration of 90 mg/ml to 110 mg/ml.

[0087] In some embodiments, said prolgolimab may be at a concentration of 100 mg/ml.

[0088] In some embodiments, said trehalose dihydrate may be present at a concentration of from 75 mg/ml to 85 mg/ml.

[0089] In some embodiments, said trehalose dihydrate may be present at a concentration of 80 mg/ml.

[0090] In some embodiments, said L-histidine may be at a concentration of 0.7 to 1.0 mg/ml.

[0091] In some embodiments, said L-histidine may be present at a concentration of 0.92 mg/ml.

[0092] In some embodiments, said L-histidine hydrochloride may be present at a concentration of 2.8 to 3.3 mg/ml.

[0093] In some embodiments, said L-histidine hydrochloride may be present at a concentration of 2.96 mg/ml.

[0094] In some embodiments, the composition may have a pH of 5.5 to 6.5.

[0095] In some embodiments, the composition may have a pH of 5.5 to 6.0.

[0096] In some embodiments, the composition may have a pH of 5.5.

[0097] The aqueous pharmaceutical composition of the anti-PD-1 antibody prolgolimab of the present invention may further contain a suitable solubilizer.

[0098] In some embodiments, said solubilizer may be poloxamer 188.

[0099] In some embodiments, said poloxamer 188 may be present in an amount that is greater than 0 mg/ml but equal to or less than 1 mg/ml.

[00100] In some embodiments, said poloxamer 188 may be present in an amount of 0 mg/ml, 0.1 mg/ml, 0.2 mg/ml, 0.3 mg/ml, 04 mg/ml, 0.5 mg/ml, 0.6 mg/ml, 0.7 mg/ml, 0.8 mg/ml, 0.9 mg/ml, 1.0 mg/ml.

[00101] In one embodiment, the present invention provides an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

(a) prolgolimab at a concentration of 20 mg/ml as an antibody;

(b) trehalose dihydrate at a concentration of 100 mg/ml;

(c) sodium acetate trihydrate at a concentration of 0.2 to 2.5 mg/ml; And

(d) acetic acid to pH 4.5 to 5.5.

[00102] In one embodiment, the present invention provides an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

(a) prolgolimab at a concentration of 20 mg/ml as an antibody;

(b) trehalose dihydrate at a concentration of 100 mg/ml;

(c) sodium acetate trihydrate at a concentration of 1.7 to 1.8 mg/ml; And

(d) acetic acid to pH 5.0.

[00103] In one embodiment, the present invention provides an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

(a) prolgolimab at a concentration of 20 mg/ml as an antibody;

(b) trehalose dihydrate at a concentration of 100 mg/ml;

(c) sodium acetate trihydrate at a concentration of 1.742 mg/ml; And

(d) acetic acid to pH 5.0.

[00104] In one embodiment, the present invention provides an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

(a) prolgolimab at a concentration of 100 mg/ml as an antibody;

(b) trehalose dihydrate at a concentration of 80 mg/ml;

(c) sodium acetate trihydrate at a concentration of 0.2 to 2.5 mg/ml; And

(d) acetic acid to pH 4.5 to 5.5.

[00105] In one embodiment, the present invention provides an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

(a) prolgolimab at a concentration of 100 mg/ml as an antibody;

(b) trehalose dihydrate at a concentration of 80 mg/ml;

(c) sodium acetate trihydrate at a concentration of 1.7 to 1.8 mg/ml; And

(d) acetic acid to a pH of 5.0 to 5.5.

[00106] In one embodiment, the present invention provides an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

(a) prolgolimab at a concentration of 100 mg/ml as an antibody;

(b) trehalose dihydrate at a concentration of 80 mg/ml;

(c) sodium acetate trihydrate at a concentration of 1.742 mg/ml; And

(d) acetic acid to a pH of 5.0 to 5.5.

[00107] In one embodiment, the present invention provides an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

(a) prolgolimab at a concentration of 100 mg/ml as an antibody;

(b) trehalose dihydrate at a concentration of 80 mg/ml;

(c) sodium acetate trihydrate at a concentration of 1.742 mg/ml; And

(d) acetic acid to pH 5.0.

[00108] In one embodiment, the present invention provides an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

(a) prolgolimab at a concentration of 20 mg/ml as an antibody;

(b) trehalose dihydrate at a concentration of 100 mg/ml;

(c) L-histidine at a concentration of 0.2 to 2.5 mg/ml; And

(d) L-histidine hydrochloride at a concentration of 0.2 to 3.5 mg/ml;

(e) wherein said composition has a pH of 5.5 to 6.5.

[00109] In one embodiment, the present invention provides an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

(a) prolgolimab at a concentration of 20 mg/ml as an antibody;

(b) trehalose dihydrate at a concentration of 100 mg/ml;

(c) L-histidine at a concentration of 0.92 mg/ml; And

(d) L-histidine hydrochloride at a concentration of 2.96 mg/ml;

(e) wherein said composition has a pH of 5.5.

[00110] In one embodiment, the present invention provides an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

(a) prolgolimab at a concentration of 100 mg/ml as an antibody;

(b) trehalose dihydrate at a concentration of 80 mg/ml;

(c) L-histidine at a concentration of 0.2 to 2.5 mg/ml; And

(d) L-histidine hydrochloride at a concentration of 0.2 to 3.5 mg/ml;

(e) wherein said composition has a pH of 5.5 to 6.5.

[00111] In one embodiment, the present invention provides an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

(a) prolgolimab at a concentration of 100 mg/ml as an antibody;

(b) trehalose dihydrate at a concentration of 80 mg/ml;

(c) L-histidine at a concentration of 0.92 mg/ml; And

(d) L-histidine hydrochloride at a concentration of 2.96 mg/ml;

(e) wherein said composition has a pH of 5.5 to 6.0.

[00112] In one embodiment, the present invention provides an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

(a) prolgolimab at a concentration of 100 mg/ml as an antibody;

(b) trehalose dihydrate at a concentration of 80 mg/ml;

(c) L-histidine at a concentration of 0.92 mg/ml; And

(d) L-histidine hydrochloride at a concentration of 2.96 mg/ml;

(e) wherein said composition has a pH of 5.5.

[00113] In one embodiment, the present invention provides an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

I. Composition per 1 ml:

Prolgolimab 2 0.0 mg Sodium acetate trihydrate 1.742 mg Trehalose dihydrate 100 mg Acetic acid ice. up to pH 5.0 Water for injections up to 1 ml.

[00114] In one embodiment, the present invention provides an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

II. Composition per 1 ml:

Prolgolimab 20.0 mg Trehalose dihydrate 100 mg L-histidine 0.92 mg L-histidine hydrochloride 2.96 mg Water for injections up to 1 ml.

[00115] The aqueous pharmaceutical composition of the anti-PD-1 antibody prolgolimab of the present invention may further contain a suitable solubilizer.

[00116] In some embodiments, said solubilizer may be poloxamer 188.

[00117] In some embodiments, said poloxamer 188 may be present in an amount that is greater than 0 mg/ml but equal to or less than 1 mg/ml.

[00118] In some embodiments, said poloxamer 188 may be present in an amount of 0 mg/ml, 0.1 mg/ml, 0.2 mg/ml, 0.3 mg/ml, 0.4 mg/ml, 0.5 mg/ml, 0.6 mg/ml, 0.7 mg/ml, 0.8 mg/ml, 0.9 mg/ml, 1.0 mg/ml.

[00119] In one embodiment, the present invention provides an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

(a) prolgolimab at a concentration of 20 mg/ml as an antibody;

(b) trehalose dihydrate at a concentration of 100 mg/ml;

(c) sodium acetate trihydrate at a concentration of 0.2 to 2.5 mg/ml; And

(d) acetic acid to pH 4.5 to 5.5;

(e) poloxamer 188 at a concentration greater than 0 mg/ml but equal to or less than 1 mg/ml.

[00120] In one embodiment, the present invention provides an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

(a) prolgolimab at a concentration of 20 mg/ml as an antibody;

(b) trehalose dihydrate at a concentration of 100 mg/ml;

(c) sodium acetate trihydrate at a concentration of 1.7 to 1.8 mg/ml; And

(d) acetic acid to pH 5.0;

(e) poloxamer 188 at a concentration greater than 0 mg/ml but equal to or less than 1 mg/ml.

[00121] In one embodiment, the present invention provides an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

(a) prolgolimab at a concentration of 20 mg/ml as an antibody;

(b) trehalose dihydrate at a concentration of 100 mg/ml;

(c) sodium acetate trihydrate at a concentration of 1.742 mg/ml; And

(d) acetic acid to pH 5.0;

(e) poloxamer 188 at a concentration greater than 0 mg/ml but equal to or less than 1 mg/ml.

[00122] In one embodiment, the present invention provides an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

(a) prolgolimab at a concentration of 100 mg/ml as an antibody;

(b) trehalose dihydrate at a concentration of 80 mg/ml;

(c) sodium acetate trihydrate at a concentration of 0.2 to 2.5 mg/ml; And

(d) acetic acid to pH 4.5 to 5.5;

(e) poloxamer 188 at a concentration greater than 0 mg/ml but equal to or less than 1 mg/ml.

[00123] In one embodiment, the present invention provides an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

(a) prolgolimab at a concentration of 100 mg/ml as an antibody;

(b) trehalose dihydrate at a concentration of 80 mg/ml;

(c) sodium acetate trihydrate at a concentration of 1.7 mg/ml to 1.8 mg/ml; And

(d) acetic acid to pH 5.0 to 5.5;

(e) poloxamer 188 at a concentration greater than 0 mg/ml but equal to or less than 1 mg/ml.

[00124] In one embodiment, the present invention provides an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

(a) prolgolimab at a concentration of 100 mg/ml as an antibody;

(b) trehalose dihydrate at a concentration of 80 mg/ml;

(c) sodium acetate trihydrate at a concentration of 1.742 mg/ml; And

(d) acetic acid to pH 5.0 to 5.5;

(e) poloxamer 188 at a concentration greater than 0 mg/ml but equal to or less than 1 mg/ml.

[00125] In one embodiment, the present invention provides an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

(a) prolgolimab at a concentration of 100 mg/ml as an antibody;

(b) trehalose dihydrate at a concentration of 80 mg/ml;

(c) sodium acetate trihydrate at a concentration of 1.742 mg/ml; And

(d) acetic acid to pH 5.0;

(e) poloxamer 188 at a concentration greater than 0 mg/ml but equal to or less than 1 mg/ml.

[00126] In one embodiment, the present invention provides an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

(a) prolgolimab at a concentration of 20 mg/ml as an antibody;

(b) trehalose dihydrate at a concentration of 100 mg/ml;

(c) L-histidine at a concentration of 0.2 to 2.5 mg/ml; And

(d) L-histidine hydrochloride at a concentration of 0.2 to 3.5 mg/ml;

(e) wherein said composition has a pH of 5.5 to 6.5;

(f) poloxamer 188 at a concentration greater than 0 mg/ml but equal to or less than 1 mg/ml.

[00127] In one embodiment, the present invention provides an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

(a) prolgolimab at a concentration of 20 mg/ml as an antibody;

(b) trehalose dihydrate at a concentration of 100 mg/ml;

(c) L-histidine at a concentration of 0.92 mg/ml; And

(d) L-histidine hydrochloride at a concentration of 2.96 mg/ml;

(e) wherein said composition has a pH of 5.5;

(f) poloxamer 188 at a concentration greater than 0 mg/ml but equal to or less than 1 mg/ml.

[00128] In one embodiment, the present invention provides an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

(a) prolgolimab at a concentration of 100 mg/ml as an antibody;

(b) trehalose dihydrate at a concentration of 80 mg/ml;

(c) L-histidine at a concentration of 0.2 to 2.5 mg/ml; And

(d) L-histidine hydrochloride at a concentration of 0.2 to 3.5 mg/ml;

(e) wherein said composition has a pH of 5.5 to 6.5;

(f) poloxamer 188 at a concentration greater than 0 mg/ml but equal to or less than 1 mg/ml.

[00129] In one embodiment, the present invention provides an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

(a) prolgolimab at a concentration of 100 mg/ml as an antibody;

(b) trehalose dihydrate at a concentration of 80 mg/ml;

(c) L-histidine at a concentration of 0.92 mg/ml; And

(d) L-histidine hydrochloride at a concentration of 2.96 mg/ml;

(e) wherein said composition has a pH of 5.5 to 6.0;

(f) poloxamer 188 at a concentration greater than 0 mg/ml but equal to or less than 1 mg/ml.

[00130] In one embodiment, the present invention provides an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

(a) prolgolimab at a concentration of 100 mg/ml as an antibody;

(b) trehalose dihydrate at a concentration of 80 mg/ml;

(c) L-histidine at a concentration of 0.92 mg/ml; And

(d) L-histidine hydrochloride at a concentration of 2.96 mg/ml;

(e) wherein said composition has a pH of 5.5;

(f) poloxamer 188 at a concentration greater than 0 mg/ml but equal to or less than 1 mg/ml.

[00131] In some embodiments of the present invention, said poloxamer 188 may be present in an amount of 0 mg/ml, 0.1 mg/ml, 0.2 mg/ml, 0.3 mg/ml, 0.4 mg/ml, 0.5 mg/ml, 0.6 mg/ml , 0.7 mg/ml, 0.8 mg/ml, 0.9 mg/ml, 1.0 mg/ml.

[00132] In some embodiments, the aqueous pharmaceutical composition of the anti-PD-1 antibody of the present invention can be administered parenterally.

[00133] In some embodiments, the aqueous pharmaceutical composition of the anti-PD-1 antibody of the present invention can be administered intramuscularly.

[00134] In some embodiments, the aqueous pharmaceutical composition of the anti-PD-1 antibody of the present invention can be administered subcutaneously.

[00135] In some embodiments, the aqueous pharmaceutical composition of the anti-PD-1 antibody of the present invention can be administered intravenously.

[00136] In some embodiments, the aqueous pharmaceutical composition of the anti-PD-1 antibody of the present invention can be administered intravenously as an infusion.

[00137] In some embodiments, the aqueous pharmaceutical composition of the anti-PD-1 antibody of the present invention can be administered intravenously as an infusion over a period of 60 minutes; if well tolerated, the infusion duration can be reduced to 30 minutes.

[00138] In one embodiment, the aqueous pharmaceutical composition of the anti-PD-1 antibody prolgolimab of the present invention may be contained in a vial.

[00139] In some embodiments, said vial may be a glass vial.

[00140] In some embodiments, said vial may have a volume of from 1 ml to 50 ml.

[00141] In some embodiments, said vial may have a volume of from 1 ml to 20 ml.

[00142] In some embodiments, said vial may have a volume of 1 ml, 2 ml, 3 ml, 4 ml, 5 ml, 6 ml, 7 ml, 8 ml, 9 ml, 10 ml, 15 ml, 20 ml, 25 ml , 30 ml, 35 ml, 4 0 ml, 4 5 ml or 5 0 ml.

[00143] In one embodiment, the aqueous pharmaceutical composition of the anti-PD-1 antibody prolgolimab of the present invention may be contained in a syringe.

[00144] In some embodiments, said syringe may have a capacity of 1 ml.

[00145] In some embodiments, said syringe may have a capacity of 2 ml.

[00146] In one embodiment, the aqueous pharmaceutical composition of the anti-PD-1 antibody prolgolimab of the present invention may be contained in a pre-filled syringe.

[00147] In some embodiments, said prefilled syringe may have a capacity of 1 ml.

[00148] In some embodiments, said prefilled syringe may have a capacity of 2 ml.

[00149] In accordance with another broad aspect of the present invention, there is provided a method for preparing an aqueous pharmaceutical composition suitable for administration to a subject to inhibit the activity of the PD-1 protein. The method includes combining a pharmaceutically effective amount of the anti-PD-1 antibody prolgolimab with an acetate-based buffering agent; and an effective amount of trehalose. The method includes combining a pharmaceutically effective amount of the anti-PD-1 antibody prolgolimab with a histidine-based buffering agent; and an effective amount of trehalose.

[00150] In some embodiments, poloxamer 188 may be added as a solubilizer.

[00151] In accordance with another broad aspect, the use of an aqueous pharmaceutical composition of the anti-PD-1 antibody prolgolimab, as defined herein, is provided for the treatment of cancer.

[00152] In some embodiments, the use of an aqueous pharmaceutical composition of the anti-PD-1 antibody prolgolimab for the treatment of malignancies, which may be selected from the group consisting of melanoma, including unresectable or metastatic melanoma, early stages of melanoma before and after curative treatment; lung cancer, non-small cell lung cancer (NSCLC), including unresectable or metastatic non-small cell lung cancer; non-squamous non-small cell lung cancer, squamous cell lung cancer; small cell lung cancer, including unresectable or metastatic small cell lung cancer; early stage lung cancer before and after radical treatment; cervical cancer, including metastatic cervical cancer, early stages of cervical cancer before and after radical treatment; head and neck tumors, including squamous cell carcinoma of the head and neck; Hodgkin's lymphoma; tumors of the stomach and intestines, metastatic squamous cell carcinoma of the esophagus; bladder cancer, including metastatic urothelial carcinoma, kidney cancer; endometrial cancer, including metastatic endometrial cancer, early stages of endometrial cancer before and after radical treatment; breast cancer, including metastatic breast cancer, early stages of endometrial cancer before and after radical treatment; liver cancer, including metastatic or inoperable liver cancer, early stages of liver cancer before and after radical treatment; unresectable or metastatic solid tumor, including unresectable or metastatic solid tumor with signs of microsatellite instability.

[00153] In another broad aspect, the use of an aqueous pharmaceutical composition of the anti-PD-1 antibody prolgolimab for the treatment of cancer is provided, comprising administering a pharmaceutically effective amount of the aqueous pharmaceutical composition as defined herein.

[00154] In another broad aspect, there is provided the use of an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

(a) prolgolimab at a concentration of 15 mg/ml to 40 mg/ml as an antibody;

(b) trehalose dihydrate at a concentration of 80 mg/ml to 110 mg/ml;

(c) sodium acetate trihydrate at a concentration of 0.2 mg/ml to 2.5 mg/ml; And

(d) acetic acid to pH 4.5-5.5,

for the treatment of malignant neoplasm in a subject in need of it.

[00155] In some embodiments, said prolgolimab may be at a concentration of 15 mg/ml to 25 mg/ml.

[00156] In some embodiments, said prolgolimab may be present at a concentration of 20 mg/ml.

[00157] In some embodiments, said trehalose dihydrate may be present at a concentration of from 95 mg/ml to 105 mg/ml.

[00158] In some embodiments, said trehalose dihydrate may be present at a concentration of 100 mg/ml.

[00159] In some embodiments, said sodium acetate trihydrate may be present at a concentration of 1.6 mg/ml to 1.9 mg/ml.

[00160] In some embodiments, said sodium acetate trihydrate may be present at a concentration of 1.7 mg/ml to 1.8 mg/ml.

[00161] In some embodiments, said sodium acetate trihydrate may be present at a concentration of 1.742 mg/ml.

[00162] In some embodiments, said acetic acid may be added to pH 5.0.

[00163] In some embodiments, said acetic acid may be present at a concentration of 0.04 to 0.77 mg/ml.

[00164] In some embodiments, said acetic acid may be present at a concentration of from 0.40 mg/ml to 0.50 mg/ml.

[00165] In some embodiments, said acetic acid may be present at a concentration of 0.4 to 3 mg/ml.

[00166] In another broad aspect, there is provided the use of an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

(a) prolgolimab at a concentration of 5 mg/ml to 40 mg/ml as an antibody;

(b) trehalose dihydrate at a concentration of 70 mg/ml to 110 mg/ml;

(c) L-histidine at a concentration of 0.2 to 2.5 mg/ml; And

(d) L-histidine hydrochloride at a concentration of 0.2 to 3.5 mg/ml; for the treatment of malignant neoplasm in a subject in need of it.

[00167] In some embodiments, said prolgolimab may be at a concentration of 15 mg/ml to 25 mg/ml.

[00168] In some embodiments, said prolgolimab may be present at a concentration of 20 mg/ml.

[00169] In some embodiments, said trehalose dihydrate may be present at a concentration of from 95 mg/ml to 105 mg/ml.

[00170] In some embodiments, said trehalose dihydrate may be present at a concentration of 100 mg/ml.

[00171] In some embodiments, said L-histidine may be present at a concentration of 0.7 to 1.0 mg/ml.

[00172] In some embodiments, said L-histidine may be present at a concentration of 0.92 mg/ml.

[00173] In some embodiments, said L-histidine hydrochloride may be present at a concentration of 2.8 to 3.3 mg/ml.

[00174] In some embodiments, said L-histidine hydrochloride may be present at a concentration of 2.96 mg/ml.

[00175] In some embodiments, the composition may have a pH of 5.5 to 6.5.

[00176] In some embodiments, the composition may have a pH of 5.5.

[00177] One embodiment of the invention provides the use of an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

(a) prolgolimab at a concentration of 20 mg/ml as an antibody;

(b) trehalose dihydrate at a concentration of 100 mg/ml;

(c) sodium acetate trihydrate at a concentration of 0.2 to 2.5 mg/ml; And

(d) acetic acid to pH 4.5 to 5.5;

for the treatment of malignant neoplasm in a subject in need of it.

[00178] One embodiment of the invention provides the use of an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

(a) prolgolimab at a concentration of 20 mg/ml as an antibody;

(b) trehalose dihydrate at a concentration of 100 mg/ml;

(c) sodium acetate trihydrate at a concentration of 1.7 mg/ml to 1.8 mg/ml; And

(d) acetic acid to pH 5.0;

for the treatment of malignant neoplasm in a subject in need of it.

[00179] One embodiment of the invention provides the use of an aqueous pharmaceutical composition of an anti-PQ-1 antibody comprising:

(a) prolgolimab at a concentration of 20 mg/ml as an antibody;

(b) trehalose dihydrate at a concentration of 100 mg/ml;

(c) sodium acetate trihydrate at a concentration of 1.742 mg/ml; And

(d) acetic acid to pH 5.0;

for the treatment of malignant neoplasm in a subject in need of it.

[00180] One embodiment of the invention provides the use of an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

(a) prolgolimab at a concentration of 20 mg/ml as an antibody;

(b) trehalose dihydrate at a concentration of 100 mg/ml;

(c) L-histidine at a concentration of 0.2 to 2.5 mg/ml; And

(d) L-histidine hydrochloride at a concentration of 0.2 to 3.5 mg/ml;

(e) wherein said composition has a pH of 5.5 to 6.5;

for the treatment of malignant neoplasm in a subject in need of it.

[00181] One embodiment of the invention provides the use of an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

(a) prolgolimab at a concentration of 20 mg/ml as an antibody;

(b) trehalose dihydrate at a concentration of 100 mg/ml;

(c) L-histidine at a concentration of 0.92 mg/ml; And

(d) L-histidine hydrochloride at a concentration of 2.96 mg/ml;

(e) wherein said composition has a pH of 5.5;

for the treatment of malignant neoplasm in a subject in need of it.

[00182] One embodiment of the invention provides the use of an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

I. Composition per 1 ml:

Prolgolimab 20.0 mg Sodium acetate trihydrate 1.742 mg Trehalose dihydrate 100 mg Acetic acid ice. up to pH 5.0 Water for injections up to 1 ml;

for the treatment of malignant neoplasm in a subject in need of it.

[00183] One embodiment of the invention provides the use of an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

II. Composition per 1 ml:

Prolgolimab 20.0 mg Trehalose dihydrate 100 mg L-histidine 0.92 mg L-histidine hydrochloride 2.96 mg Water for injections up to 1 ml;

for the treatment of malignant neoplasm in a subject in need of it.

[00184] The aqueous pharmaceutical composition of the anti-PD-1 antibody prolgolimab of the present invention may further contain a suitable solubilizer.

[00185] In some embodiments, said solubilizer may be poloxamer 188.

[00186] In some embodiments, said poloxamer 188 may be present in an amount that is greater than 0 mg/ml but equal to or less than 1 mg/ml.

[00187] In some embodiments, said poloxamer 188 may be present in an amount of 0 mg/ml, 0.1 mg/ml, 0.2 mg/ml, 0.3 mg/ml, 04 mg/ml, 0.5 mg/ml, 0.6 mg/ml, 0.7 mg/ml, 0.8 mg/ml, 0.9 mg/ml, 1.0 mg/ml.

[00188] In some embodiments, use of said aqueous pharmaceutical composition of the anti-PD-1 antibody prolgolimab may comprise administering the composition at a dose of the anti-PD-1 antibody prolgolimab of 1 mg/kg body weight.

[00189] In some embodiments, use of said aqueous pharmaceutical composition of the anti-PD-1 antibody prolgolimab may include administering the composition at a dose of 3 mg/kg body weight of the anti-PD-1 antibody prolgolimab.

[00190] In some embodiments, use of said aqueous pharmaceutical composition of the anti-PD-1 antibody prolgolimab may include administering the composition every 2 weeks.

[00191] In some embodiments, use of said aqueous pharmaceutical composition of the anti-PD-1 antibody prolgolimab may include administering the composition once every 2 weeks.

[00192] In some embodiments, use of said aqueous pharmaceutical composition of the anti-PD-1 antibody prolgolimab may include administering the composition every 3 weeks.

[00193] In some embodiments, use of said aqueous pharmaceutical composition of the anti-PD-1 antibody prolgolimab may include administering the composition once every 3 weeks.

[00194] In some embodiments, use of said aqueous pharmaceutical composition of the anti-PD-1 antibody prolgolimab may include administering the composition at a dose of the anti-PD-1 antibody prolgolimab of 1 mg/kg body weight every 2 weeks.

[00195] In some embodiments, use of said aqueous pharmaceutical composition of the anti-PD-1 antibody prolgolimab may include administering the composition at a dose of the anti-PD-1 antibody prolgolimab of 1 mg/kg body weight once every 2 weeks.

[00196] In some embodiments, use of said aqueous pharmaceutical composition of the anti-PD-1 antibody prolgolimab may include administering the composition at a dose of the anti-PD-1 antibody prolgolimab of 3 mg/kg body weight every 3 weeks.

[00197] In some embodiments, use of said aqueous pharmaceutical composition of the anti-PD-1 antibody prolgolimab may include administering the composition at a dose of the anti-PD-1 antibody prolgolimab of 3 mg/kg body weight once every 3 weeks.

[00198] In some embodiments, use of said aqueous pharmaceutical composition of the anti-PD-1 antibody prolgolimab may include administering the composition parenterally.

[00199] In some embodiments, said parenteral administration may be intravenous, subcutaneous, or intramuscular administration.

[00200] In some embodiments, use of said aqueous pharmaceutical composition of the anti-PD-1 antibody prolgolimab may include administering the composition intravenously as an infusion.

[00201] In some embodiments, the aqueous pharmaceutical composition of the anti-PD-1 antibody prolgolimab of the present invention may be administered intravenously as a 60-minute infusion, which may be reduced to 30 minutes if well tolerated.

[00202] In some embodiments, the malignancy is melanoma, including unresectable or metastatic melanoma, early stages of melanoma before and after curative treatment; lung cancer, non-small cell lung cancer (NSCLC), including unresectable or metastatic non-small cell lung cancer.

[00203] In one embodiment, the invention provides a method of treating cancer in a subject in need thereof, comprising administering a therapeutically effective amount of an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

(a) prolgolimab at a concentration of 15 mg/ml to 40 mg/ml as an antibody;

(b) trehalose dihydrate at a concentration of 80 mg/ml to 110 mg/ml;

(c) sodium acetate trihydrate at a concentration of 0.2 mg/ml to 2.5 mg/ml; And

(d) acetic acid to pH 4.5-5.5.

[00204] In some embodiments, said prolgolimab may be at a concentration of 15 mg/ml to 25 mg/ml.

[00205] In some embodiments, said prolgolimab may be at a concentration of 20 mg/ml.

[00206] In some embodiments, said trehalose dihydrate may be present at a concentration of from 95 mg/ml to 105 mg/ml.

[00207] In some embodiments, said trehalose dihydrate may be present at a concentration of 100 mg/ml.

[00208] In some embodiments, said sodium acetate trihydrate may be present at a concentration of 1.6 mg/ml to 1.9 mg/ml.

[00209] In some embodiments, said sodium acetate trihydrate may be present at a concentration of 1.7 mg/ml to 1.8 mg/ml.

[00210] In some embodiments, said sodium acetate trihydrate may be present at a concentration of 1.742 mg/ml.

[00211] In some embodiments, said acetic acid may be added to pH 5.0.

[00212] In some embodiments, said acetic acid may be present at a concentration of 0.04 to 0.77 mg/ml.

[00213] In some embodiments, said acetic acid may be present at a concentration of 0.40 mg/ml to 0.50 mg/ml.

[00214] In some embodiments, said acetic acid may be present at a concentration of 0.4 to 3 mg/ml.

[00215] In one embodiment, the invention provides a method of treating cancer in a subject in need thereof, comprising administering a therapeutically effective amount of an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

(a) prolgolimab at a concentration of 5 mg/ml to 40 mg/ml as an antibody;

(b) trehalose dihydrate at a concentration of 70 mg/ml to 110 mg/ml;

(c) L-histidine at a concentration of 0.2 to 2.5 mg/ml; And

(d) L-histidine hydrochloride at a concentration of 0.2 to 3.5 mg/ml.

[00216] In some embodiments, said prolgolimab may be at a concentration of 15 mg/ml to 25 mg/ml.

[00217] In some embodiments, said prolgolimab may be at a concentration of 20 mg/ml.

[00218] In some embodiments, said trehalose dihydrate may be at a concentration of from 95 mg/ml to 105 mg/ml.

[00219] In some embodiments, said trehalose dihydrate may be present at a concentration of 100 mg/ml.

[00220] In some embodiments, said L-histidine may be at a concentration of 0.7 to 1.0 mg/ml.

[00221] In some embodiments, said L-histidine may be present at a concentration of 0.92 mg/ml.

[00222] In some embodiments, said L-histidine hydrochloride may be present at a concentration of 2.8 to 3.3 mg/ml.

[00223] In some embodiments, said L-histidine hydrochloride may be present at a concentration of 2.96 mg/ml.

[00224] In some embodiments, the composition may have a pH of 5.5 to 6.5.

[00225] In some embodiments, the composition may have a pH of 5.5.

[00226] In one embodiment, the invention provides a method of treating cancer in a subject in need thereof, comprising administering a therapeutically effective amount of an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

(a) prolgolimab at a concentration of 20 mg/ml as an antibody;

(b) trehalose dihydrate at a concentration of 100 mg/ml;

(c) sodium acetate trihydrate at a concentration of 0.2 to 2.5 mg/ml; And

(d) acetic acid to pH 4.5 to 5.5.

[00227] In one embodiment, the invention provides a method of treating cancer in a subject in need thereof, comprising administering a therapeutically effective amount of an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

(a) prolgolimab at a concentration of 20 mg/ml as an antibody;

(b) trehalose dihydrate at a concentration of 100 mg/ml;

(c) sodium acetate trihydrate at a concentration of 1.7 mg/ml to 1.8 mg/ml; And

(d) acetic acid to pH 5.0.

[00228] In one embodiment, the invention provides a method of treating cancer in a subject in need thereof, comprising administering a therapeutically effective amount of an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

(a) prolgolimab at a concentration of 20 mg/ml as an antibody;

(b) trehalose dihydrate at a concentration of 100 mg/ml;

(c) sodium acetate trihydrate at a concentration of 1.742 mg/ml; And

(d) acetic acid to pH 5.0.

[00229] In one embodiment, the invention provides a method of treating cancer in a subject in need thereof, comprising administering a therapeutically effective amount of an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

(a) prolgolimab at a concentration of 20 mg/ml as an antibody;

(b) trehalose dihydrate at a concentration of 100 mg/ml;

(c) L-histidine at a concentration of 0.2 to 2.5 mg/ml; And

(d) L-histidine hydrochloride at a concentration of 0.2 to 3.5 mg/ml;

(e) wherein said composition has a pH of 5.5 to 6.5.

[00230] In one embodiment, the invention provides a method of treating cancer in a subject in need thereof, comprising administering a therapeutically effective amount of an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

(a) prolgolimab at a concentration of 20 mg/ml as an antibody;

(b) trehalose dihydrate at a concentration of 100 mg/ml;

(c) L-histidine at a concentration of 0.92 mg/ml; And

(d) L-histidine hydrochloride at a concentration of 2.96 mg/ml;

(e) wherein said composition has a pH of 5.5.

[00231] In one embodiment, the invention provides a method of treating cancer in a subject in need thereof, comprising administering a therapeutically effective amount of an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

I. Composition per 1 ml:

Prolgolimab 20.0 mg Sodium acetate trihydrate 1.742 mg Trehalose dihydrate 100 mg Acetic acid ice. up to pH 5.0 Water for injections up to 1 ml.

[00232] In one embodiment, the invention provides a method of treating cancer in a subject in need thereof, comprising administering a therapeutically effective amount of an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

II. Composition per 1 ml:

Prolgolimab 20.0 mg Trehalose dihydrate 100 mg L-histidine 0.92 mg L-histidine hydrochloride 2.96 mg Water for injections up to 1 ml.

[00233] The aqueous pharmaceutical composition of the anti-PD-1 antibody prolgolimab of the present invention may further contain a suitable solubilizer.

[00234] In some embodiments, said solubilizer may be poloxamer 188.

[00235] In some embodiments, said poloxamer 188 may be present in an amount that is greater than 0 mg/ml but equal to or less than 1 mg/ml.

[00236] In some embodiments, said poloxamer 188 may be present in an amount of 0 mg/ml, 0.1 mg/ml, 0.2 mg/ml, 0.3 mg/ml, 0.4 mg/ml, 0.5 mg/ml, 0.6 mg/ml, 0.7 mg/ml, 0.8 mg/ml, 0.9 mg/ml, 1.0 mg/ml.

[00237] In some embodiments, a method of treating cancer in a subject in need thereof may comprise administering a therapeutically effective amount of said aqueous pharmaceutical composition of the anti-PD-1 antibody prolgolimab at a dose of the anti-PD-1 antibody prolgolimab of 1 mg/kg body weight.

[00238] In some embodiments, a method of treating cancer in a subject in need thereof may comprise administering a therapeutically effective amount of said aqueous pharmaceutical composition of the anti-PD-1 antibody prolgolimab at a dose of the anti-PD-1 antibody prolgolimab of 3 mg/kg body weight.

[00239] In some embodiments, a method of treating cancer in a subject in need thereof may comprise administering a therapeutically effective amount of said aqueous pharmaceutical composition of the anti-PD-1 antibody prolgolimab every 2 weeks.

[00240] In some embodiments, a method of treating cancer in a subject in need thereof may comprise administering a therapeutically effective amount of said aqueous pharmaceutical composition of the anti-PD-1 antibody prolgolimab once every 2 weeks.

[00241] In some embodiments, a method of treating cancer in a subject in need thereof may comprise administering a therapeutically effective amount of said aqueous pharmaceutical composition of the anti-PD-1 antibody prolgolimab every 3 weeks.

[00242] In some embodiments, a method of treating cancer in a subject in need thereof may comprise administering a therapeutically effective amount of said aqueous pharmaceutical composition of the anti-PD-1 antibody prolgolimab once every 3 weeks.

[00243] In some embodiments, a method of treating cancer in a subject in need thereof may comprise administering a therapeutically effective amount of said aqueous pharmaceutical composition of the anti-PD-1 antibody prolgolimab at a dose of the anti-PD-1 antibody prolgolimab of 1 mg/kg body weight every 2 weeks. .

[00244] In some embodiments, a method of treating cancer in a subject in need thereof may comprise administering a therapeutically effective amount of said aqueous pharmaceutical composition of the anti-PD-1 antibody prolgolimab at a dose of the anti-PD-1 antibody prolgolimab 1 mg/kg body weight once per 2 weeks.

[00245] In some embodiments, a method of treating cancer in a subject in need thereof may comprise administering a therapeutically effective amount of said aqueous pharmaceutical composition of the anti-PD-1 antibody prolgolimab at a dose of the anti-PD-1 antibody prolgolimab of 3 mg/kg body weight every 3 weeks. .

[00246] In some embodiments, a method of treating cancer in a subject in need thereof may comprise administering a therapeutically effective amount of said aqueous pharmaceutical composition of the anti-PD-1 antibody prolgolimab at a dose of the anti-PD-1 antibody prolgolimab 3 mg/kg body weight once per 3 weeks.

[00247] In some embodiments, a method of treating cancer in a subject in need thereof may comprise administering a therapeutically effective amount of said aqueous pharmaceutical composition of the anti-PD-1 antibody prolgolimab parenterally.

[00248] In some embodiments, said parenteral administration may be intravenous, subcutaneous, or intramuscular administration.

[00249] In some embodiments, a method of treating cancer in a subject in need thereof may comprise administering a therapeutically effective amount of said aqueous pharmaceutical composition of the anti-PD-1 antibody prolgolimab intravenously as an infusion.

[00250] In some embodiments, the aqueous pharmaceutical composition of the anti-PD-1 antibody prolgolimab of the present invention may be administered intravenously as a 60-minute infusion, which may be reduced to 30 minutes if tolerated.

[00251] In some embodiments, the malignancy is melanoma, including unresectable or metastatic melanoma, early stages of melanoma before and after curative treatment; lung cancer, non-small cell lung cancer (NSCLC), including unresectable or metastatic non-small cell lung cancer.

[00252] In one embodiment, the invention provides a method of treating cancer in a subject in need thereof, comprising administering a therapeutically effective amount of prolgolimab.

[00253] In some embodiments, a method of treating cancer in a subject in need thereof may comprise administering a therapeutically effective amount of prolgolimab at a dose of 1 mg/kg.

[00254] In some embodiments, a method of treating cancer in a subject in need thereof may comprise administering a therapeutically effective amount of prolgolimab at a dose of 3 mg/kg.

[00255] In some embodiments, a method of treating cancer in a subject in need thereof may comprise administering a therapeutically effective amount of prolgolimab every 2 weeks.

[00256] In some embodiments, a method of treating cancer in a subject in need thereof may comprise administering a therapeutically effective amount of prolgolimab once every 2 weeks.

[00257] In some embodiments, a method of treating cancer in a subject in need thereof may comprise administering a therapeutically effective amount of prolgolimab every 3 weeks.

[00258] In some embodiments, a method of treating cancer in a subject in need thereof may comprise administering a therapeutically effective amount of prolgolimab once every 3 weeks.

[00259] In some embodiments, a method of treating cancer in a subject in need thereof may comprise administering a therapeutically effective amount of prolgolimab at a dose of 1 mg/kg every 2 weeks.

[00260] In some embodiments, a method of treating cancer in a subject in need thereof may comprise administering a therapeutically effective amount of prolgolimab at a dose of 1 mg/kg once every 2 weeks.

[00261] In some embodiments, a method of treating cancer in a subject in need thereof may comprise administering a therapeutically effective amount of prolgolimab at a dose of 3 mg/kg every 3 weeks.

[00262] In some embodiments, a method of treating cancer in a subject in need thereof may comprise administering a therapeutically effective amount of prolgolimab at a dose of 3 mg/kg once every 3 weeks.

[00263] In some embodiments, a method of treating cancer in a subject in need thereof may comprise administering a therapeutically effective amount of prolgolimab parenterally.

[00264] In some embodiments, said parenteral administration may be intravenous, subcutaneous, or intramuscular administration.

[00265] In some embodiments, a method of treating cancer in a subject in need thereof may comprise administering a therapeutically effective amount of prolgolimab intravenously as an infusion.

[00266] In some embodiments, prolgolimab may be administered intravenously as a 60-minute infusion, which may be reduced to 30 minutes if tolerated.

[00267] In some embodiments, the malignancy is melanoma, including unresectable or metastatic melanoma, early stages of melanoma before and after curative treatment; lung cancer, non-small cell lung cancer (NSCLC), including unresectable or metastatic non-small cell lung cancer.

[00268] In one embodiment, the invention provides an aqueous pharmaceutical composition suitable for administration to a subject to inhibit the activity of the PD-1 protein, wherein the pharmaceutical composition contains, for every 1 ml of the pharmaceutical composition:

Prolgolimab 20.0 mg Sodium acetate trihydrate 1.742 mg Trehalose dihydrate 100 mg Acetic acid ice. up to pH 5.0 Water for injections up to 1 ml.

[00269] In one embodiment, the invention provides an aqueous pharmaceutical composition suitable for administration to a subject to inhibit the activity of the PD-1 protein, wherein the pharmaceutical composition contains for every 1 ml of the pharmaceutical composition:

Prolgolimab 20.0 mg Trehalose dihydrate 100 mg L-histidine 0.92 mg L-histidine hydrochloride 2.96 mg Water for injections up to 1 ml.

[00270] In some embodiments, the aqueous pharmaceutical composition suitable for administration to a subject to inhibit PD-1 protein activity may be administered at a dose of the anti-PD-1 antibody prolgolimab of 1 mg/kg body weight.

[00271] In some embodiments, the aqueous pharmaceutical composition suitable for administration to a subject to inhibit PD-1 protein activity may be administered at a dose of the anti-PD-1 antibody prolgolimab of 3 mg/kg body weight.

[00272] In some embodiments, the aqueous pharmaceutical composition suitable for administration to a subject to inhibit PD-1 protein activity may be administered every 2 weeks.

[00273] In some embodiments, the aqueous pharmaceutical composition suitable for administration to a subject to inhibit PD-1 protein activity may be administered once every 2 weeks.

[00274] In some embodiments, the aqueous pharmaceutical composition suitable for administration to a subject to inhibit PD-1 protein activity may be administered once every 3 weeks.

[00275] In some embodiments, the aqueous pharmaceutical composition suitable for administration to a subject to inhibit PD-1 protein activity may be administered at a dose of the anti-PD-1 antibody prolgolimab of 1 mg/kg body weight every 2 weeks.

[00276] In some embodiments, the aqueous pharmaceutical composition suitable for administration to a subject to inhibit PD-1 protein activity may be administered at a dose of the anti-PD-1 antibody prolgolimab of 1 mg/kg body weight once every 2 weeks.

[00277] In some embodiments, the aqueous pharmaceutical composition suitable for administration to a subject to inhibit PD-1 protein activity may be administered at a dose of the anti-PD-1 antibody prolgolimab of 3 mg/kg body weight every 3 weeks.

[00278] In some embodiments, the aqueous pharmaceutical composition suitable for administration to a subject to inhibit PD-1 protein activity may be administered at a dose of the anti-PD-1 antibody prolgolimab of 3 mg/kg body weight once every 3 weeks.

[00279] In some embodiments, a method of treating cancer in a subject in need thereof may comprise administering a therapeutically effective amount of prolgolimab parenterally.

[00280] In some embodiments, said parenteral administration may be intravenous, subcutaneous, or intramuscular.

[00281] In some embodiments, a method of treating cancer in a subject in need thereof may comprise administering a therapeutically effective amount of prolgolimab intravenously as an infusion.

[00282] In some embodiments, prolgolimab may be administered intravenously as a 60-minute infusion, which may be reduced to 30 minutes if tolerated.

[00283] In some embodiments, the malignancy is melanoma, including unresectable or metastatic melanoma, early stages of melanoma before and after curative treatment; lung cancer, non-small cell lung cancer (NSCLC), including unresectable or metastatic non-small cell lung cancer.

EXAMPLE IMPLEMENTATION OPTIONS:

[00284] The present invention relates to suitable aqueous pharmaceutical compositions of the anti-PD-1 antibody prolgolimab. In one embodiment of the invention, the aqueous pharmaceutical composition of prolgolimab may contain an acetate-based buffer and trehalose. Poloxamer 188 can be added as a solubilizer. In yet another embodiment, the aqueous pharmaceutical composition of prolgolimab may contain a histidine-based buffer and trehalose. Poloxamer 188 can be added as a solubilizer.

[00285] A histidine-based buffer can be formed by combining L-histidine with histidine hydrochloride or, optionally, with hydrochloric acid or other acids. It should be understood that while histidine hydrochloride can be used as the salt for the histidine buffer, any other histidine salt can be used for the histidine buffer without departing from the teachings of the present invention.

[00286] An acetate-based buffer can be formed by combining acetic acid with sodium acetate trihydrate. It should be understood that while sodium acetate trihydrate can be used as the salt for the acetate buffer, any other acetate salt, such as potassium acetate, can be used for the acetate buffer without departing from the teachings of the present invention.

[00287] In addition, the composition of the present invention may further include one or more other suitable excipients that are well known to those skilled in the art.

[00288] In some embodiments, the liquid pharmaceutical composition is formulated such that it is shelf stable in the sense that no further protein aggregation or modification occurs beyond the stability at zero time point.

[00289] In one embodiment, the inventors have unexpectedly produced highly concentrated aqueous pharmaceutical compositions of the anti-PD-1 antibody prolgolimab, where prolgolimab can be at a concentration of from 90 mg/ml to 150 mg/ml. In some embodiments of the present invention, the specified prolgolimab may be at a concentration of 90 mg/ml, 95 mg/ml, 100 mg/ml, 105 mg/ml, 110 mg/ml, 115 mg/ml, 120 mg/ml, 125 mg/ml ml, 130 mg/ml, 135 mg/ml, 140 mg/ml, 145 mg/ml, 150 mg/ml.

[00290] Such highly concentrated aqueous pharmaceutical compositions of the anti-PD-1 antibody prolgolimab of the present invention showed colloidal stability under vigorous stirring (800 rpm) for 120 hours and high thermal stability when heated at 50°C and 37°C, and also an acceptable viscosity for parenteral administration is less than 50 cP.

[00291] The above compositions are suitable for parenteral administration, such as intravenous, subcutaneous, intradermal, intraarterial, intrathecal, intraperitoneal, intraarticular and/or intramuscular administration.

[00292] The provided pharmaceutical compositions can be administered to an individual in need of treatment by systemic injection, for example, by intravenous or subcutaneous or intramuscular injection; or by injection or application to the appropriate site, for example, by direct injection or direct application to the site when the site is accessible by surgery; or through topical application.

[00293] The above compositions can be administered to an individual in need of treatment intravenously as an infusion.

[00294] In some embodiments, the aqueous pharmaceutical composition of the anti-PD-1 antibody prolgolimab of the present invention may be administered intravenously as a 60-minute infusion, which may be reduced to 30 minutes if well tolerated.

[00295] The aqueous pharmaceutical composition of the anti-PD-1 antibody prolgolimab of the present invention can be used after dilution. To do this, the required amount of the composition from the bottle is transferred into an infusion container containing a sterile 0.9% sodium chloride solution or a sterile 5% dextrose solution. The concentration of this composition in the prepared solution can range from 0.5 to 10 mg/ml. The prepared solution is mixed by carefully inverting the infusion container to avoid foaming.

Methods of treatment and use of aqueous composition

[00296] In another embodiment, the present invention provides a method of treating a mammal, comprising administering to the mammal a therapeutically effective amount of pharmaceutical compositions, wherein the concentration of the anti-PD-1 antibody prolgolimab is from 15 mg/ml to 40 mg/ml, according to the present invention, wherein the mammal may have a disease or disorder that can be effectively treated with the anti-PD-1 antibody prolgolimab of the present invention.

[00297] In another embodiment, the present invention provides a method of treating a mammal, comprising administering to the mammal a therapeutically effective amount of pharmaceutical compositions wherein the concentration of the anti-PD-1 antibody prolgolimab is 20 mg/ml, according to the present invention, wherein the mammal may have a disease or disorder , which can be effectively treated with the anti-PD-1 antibody prolgolimab of the present invention.

[00298] In a preferred embodiment, the mammal is a human.

[00299] In another embodiment, the present invention provides a method of treating a subject in need thereof, comprising administering to the subject a therapeutically effective amount of pharmaceutical compositions wherein the concentration of the anti-PD-1 antibody prolgolimab is from 15 mg/ml to 40 mg/ml, according to the present invention wherein the subject may have a disease or disorder that can be effectively treated with the anti-PD-1 antibody prolgolimab of the present invention.

[00300] In another embodiment, the present invention provides a method of treating a subject in need thereof, comprising administering to the subject a therapeutically effective amount of pharmaceutical compositions wherein the concentration of the anti-PD-1 antibody prolgolimab is 20 mg/ml, according to the present invention, wherein the subject may have a disease or disorder that can be effectively treated with the anti-PD-1 antibody prolgolimab of the present invention.

[00301] In a preferred embodiment, the subject is a human.

[00302] In one embodiment, the invention relates to a method of treating melanoma, comprising administering to a subject in need thereof a therapeutically effective amount of one of the following compositions, wherein the concentration of the anti-PD-1 antibody prolgolimab is from 15 mg/ml to 40 mg/ml, according to this invention.

[00303] In one embodiment, the invention provides a method of treating melanoma, comprising administering to a subject in need thereof a therapeutically effective amount of one of the following compositions, wherein the concentration of the anti-PD-1 antibody prolgolimab is 20 mg/ml, of the invention.

[00304] In one embodiment, the invention provides a method of treating inoperable melanoma, comprising administering to a subject in need thereof a therapeutically effective amount of one of the following compositions of the anti-PD-1 antibody prolgolimab, wherein the concentration of the anti-PD-1 antibody prolgolimab is between 15 mg/ml up to 40 mg/ml, according to this invention.

[00305] In one embodiment, the invention provides a method of treating inoperable melanoma, comprising administering to a subject in need thereof a therapeutically effective amount of one of the following compositions of the anti-PD-1 antibody prolgolimab, wherein the concentration of the anti-PD-1 antibody prolgolimab is 20 mg/ml, according to this invention.

[00306] In one embodiment, the invention provides a method of treating metastatic melanoma, comprising administering to a subject in need thereof a therapeutically effective amount of one of the following compositions of the anti-PD-1 antibody prolgolimab, wherein the concentration of the anti-PD-1 antibody prolgolimab is between 15 mg/ml up to 40 mg/ml, according to this invention.

[00307] In one embodiment, the invention provides a method of treating metastatic melanoma, comprising administering to a subject in need thereof a therapeutically effective amount of one of the following compositions of the anti-PD-1 antibody prolgolimab, wherein the concentration of the anti-PD-1 antibody prolgolimab is 20 mg/ml , according to this invention.

[00308] In one embodiment, the invention provides a method of treating early stages of melanoma before and after curative treatment, comprising administering to a subject in need thereof a therapeutically effective amount of one of the following compositions of the anti-PD-1 antibody prolgolimab, wherein the concentration of the anti-PD-1 antibody prolgolimab ranges from 15 mg/ml to 40 mg/ml, according to this invention.

[00309] In one embodiment, the invention provides a method of treating early stages of melanoma before and after curative treatment, comprising administering to a subject in need thereof a therapeutically effective amount of one of the following compositions of the anti-PD-1 antibody prolgolimab, wherein the concentration of the anti-PD-1 antibody prolgolimab is 20 mg/ml, according to this invention.

[00310] In one embodiment, the invention provides a method of treating lung cancer, comprising administering to a subject in need thereof a therapeutically effective amount of one of the following compositions of the anti-PD-1 antibody prolgolimab, wherein the concentration of the anti-PD-1 antibody prolgolimab is between 15 mg/ml up to 40 mg/ml, according to this invention.

[00311] In one embodiment, the invention provides a method of treating lung cancer, comprising administering to a subject in need thereof a therapeutically effective amount of one of the provided anti-PD-1 antibody prolgolimab compositions, wherein the concentration of the anti-PD-1 antibody prolgolimab is 20 mg/ml, according to this invention.

[00312] In one embodiment, the invention provides a method of treating non-small cell lung cancer (NSCLC), comprising administering to a subject in need thereof a therapeutically effective amount of one of the provided compositions of the anti-PD-1 antibody prolgolimab, wherein the concentration of the anti-PD-1 antibody prolgolimab is from 15 mg/ml to 40 mg/ml, according to this invention.

[00313] In one embodiment, the invention provides a method of treating non-small cell lung cancer (NSCLC), comprising administering to a subject in need thereof a therapeutically effective amount of one of the provided compositions of the anti-PD-1 antibody prolgolimab, wherein the concentration of the anti-PD-1 antibody prolgolimab is 20 mg/ml, according to this invention.

[00314] In one embodiment, the invention provides a method of treating unresectable or metastatic non-small cell lung cancer, comprising administering to a subject in need thereof a therapeutically effective amount of one of the provided anti-PD-1 antibody prolgolimab compositions, wherein the concentration of the anti-PD-1 antibody prolgolimab is from 15 mg/ml to 40 mg/ml, according to this invention.

[00315] In one embodiment, the invention provides a method of treating unresectable or metastatic non-small cell lung cancer, comprising administering to a subject in need thereof a therapeutically effective amount of one of the provided anti-PD-1 antibody prolgolimab compositions, wherein the concentration of the anti-PD-1 antibody prolgolimab is 20 mg/ml, according to this invention.

[00316] In one embodiment, the invention provides a method of treating melanoma in a subject in need thereof, comprising administering a therapeutically effective amount of an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

I. Composition per 1 ml:

Prolgolimab 20.0 mg Sodium acetate trihydrate 1.742 mg Trehalose dihydrate 100 mg Acetic acid ice. up to pH 5.0 Water for injections up to 1 ml.

[00317] In one embodiment, the invention provides a method of treating melanoma in a subject in need thereof, comprising administering a therapeutically effective amount of an aqueous anti-PD-1 antibody pharmaceutical composition comprising:

II. Composition per 1 ml:

Prolgolimab 20.0 mg Trehalose dihydrate 100 mg L-histidine 0.92 mg L-histidine hydrochloride 2.9 6 mg Water for injections up to 1 ml.

[00318] In one embodiment, the invention provides a method of treating unresectable melanoma in a subject in need thereof, comprising administering a therapeutically effective amount of an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

I. Composition per 1 ml:

Prolgolimab 20.0 mg Sodium acetate trihydrate 1.742 mg Trehalose dihydrate 100 mg Acetic acid ice. up to pH 5.0 Water for injections up to 1 ml.

[00319] In one embodiment, the invention provides a method of treating unresectable melanoma in a subject in need thereof, comprising administering a therapeutically effective amount of an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

II. Composition per 1 ml:

Prolgolimab 20.0 mg Trehalose dihydrate 100 mg L-histidine 0.92 mg L-histidine hydrochloride 2.96 mg Water for injections up to 1 ml.

[00320] In one embodiment, the invention provides a method of treating metastatic melanoma in a subject in need thereof, comprising administering a therapeutically effective amount of an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

I. Composition per 1 ml:

Prolgolimab 20.0 mg Sodium acetate trihydrate 1.742 mg Trehalose dihydrate 100 mg Acetic acid ice. up to pH 5.0 Water for injections up to 1 ml.

[00321] In one embodiment, the invention provides a method of treating metastatic melanoma in a subject in need thereof, comprising administering a therapeutically effective amount of an aqueous anti-PD-1 antibody pharmaceutical composition comprising:

II. Composition per 1 ml:

Prolgolimab 20.0 mg Trehalose dihydrate 100 mg L-histidine 0.92 mg L-histidine hydrochloride 2.96 mg Water for injections up to 1 ml.

[00322] In one embodiment, the invention provides a method of treating early stages of melanoma before and after curative treatment in a subject in need thereof, comprising administering a therapeutically effective amount of an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

I. Composition per 1 ml:

Prolgolimab 20.0 mg Sodium acetate trihydrate 1.742 mg Trehalose dihydrate 100 mg Acetic acid ice. up to pH 5.0 Water for injections up to 1 ml.

[00323] In one embodiment, the invention provides a method of treating early stages of melanoma before and after curative treatment in a subject in need thereof, comprising administering a therapeutically effective amount of an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

II. Composition per 1 ml:

Prolgolimab 20.0 mg Trehalose dihydrate 100 mg L-histidine 0.92 mg L-histidine hydrochloride 2.96 mg Water for injections up to 1 ml.

[00324] In one embodiment, the invention provides a method of treating lung cancer in a subject in need thereof, comprising administering a therapeutically effective amount of aqueous

a pharmaceutical composition of an anti-PD-1 antibody containing:

I. Composition per 1 ml:

Prolgolimab 20.0 mg Sodium acetate trihydrate 1.742 mg Trehalose dihydrate 100 mg Acetic acid ice. up to pH 5.0 Water for injections up to 1 ml.

[00325] In one embodiment, the invention provides a method of treating lung cancer in a subject in need thereof, comprising administering a therapeutically effective amount of an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

II. Composition per 1 ml:

Prolgolimab 20.0 mg Trehalose dihydrate 100 mg L-histidine 0.92 mg L-histidine hydrochloride 2.96 mg Water for injections up to 1 ml.

[00326] In one embodiment, the invention provides a method of treating non-small cell lung cancer (NSCLC) in a subject in need thereof, comprising administering a therapeutically effective amount of an aqueous anti-PD-1 antibody pharmaceutical composition comprising:

I. Composition per 1 ml:

Prolgolimab 20.0 mg Sodium acetate trihydrate 1.742 mg Trehalose dihydrate 100 mg Acetic acid ice. up to pH 5.0 Water for injections up to 1 ml.

[00327] In one embodiment, the invention provides a method of treating non-small cell lung cancer (NSCLC) in a subject in need thereof, comprising administering a therapeutically effective amount of an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

II. Composition per 1 ml:

Prolgolimab 20.0 mg Trehalose dihydrate 100 mg L-histidine 0.92 mg L-histidine hydrochloride 2.96 mg Water for injections up to 1 ml.

[00328] In one embodiment, the invention provides a method of treating unresectable or metastatic non-small cell lung cancer in a subject in need thereof, comprising administering a therapeutically effective amount of an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

I. Composition per 1 ml:

Prolgolimab 20.0 mg Sodium acetate trihydrate 1.742 mg Trehalose dihydrate 100 mg Acetic acid ice. up to pH 5.0 Water for injections up to 1 ml.

[00329] In one embodiment, the invention provides a method of treating unresectable or metastatic non-small cell lung cancer in a subject in need thereof, comprising administering a therapeutically effective amount of an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

II. Composition per 1 ml:

Prolgolimab 20.0 mg Trehalose dihydrate 100 mg L-histidine 0.92 mg L-histidine hydrochloride 2.96 mg Water for injections up to 1 ml.

[00330] In one embodiment, the invention provides a method of treating melanoma, comprising administering to a subject in need thereof a therapeutically effective amount of the anti-PD-1 antibody prolgolimab.

[00331] In one embodiment, the invention provides a method of treating unresectable melanoma, comprising administering to a subject in need thereof a therapeutically effective amount of the anti-PD-1 antibody prolgolimab.

[00332] In one embodiment, the invention provides a method of treating metastatic melanoma, comprising administering to a subject in need thereof a therapeutically effective amount of the anti-PD-1 antibody prolgolimab.

[00333] In one embodiment, the invention provides a method of treating early stages of melanoma before and after curative treatment, comprising administering to a subject in need thereof a therapeutically effective amount of the anti-PD-1 antibody prolgolimab.

[00334] In one embodiment, the invention provides a method of treating lung cancer, comprising administering to a subject in need thereof a therapeutically effective amount of the anti-PD-1 antibody prolgolimab.

[00335] In one embodiment, the invention provides a method of treating non-small cell lung cancer (NSCLC), comprising administering to a subject in need thereof a therapeutically effective amount of the anti-PD-1 antibody prolgolimab.

[00336] In one embodiment, the invention provides a method of treating unresectable or metastatic non-small cell lung cancer, comprising administering to a subject in need thereof a therapeutically effective amount of the anti-PD-1 antibody prolgolimab.

[00337] The therapeutically effective amount of the anti-PD-1 antibody prolgolimab of the present invention, and aqueous compositions comprising the anti-PD-1 antibody prolgolimab of the present invention, in the present formulations depends on the condition being treated, the severity of the condition, prior therapy and medical history patient and response to therapeutic agent. The appropriate dose can be adjusted according to the discretion of the attending physician so that it can be administered to the patient once or through multiple administrations.

[00338] In one embodiment, the effective amount of anti-PD-1 antibody per dose to a patient is about 0.01 to 10 mg per kilogram of body weight, or about 1 to 10 mg per kilogram of body weight, or about 0.05 mg per kilogram of body weight body weight, or approximately 0.25 mg per kilogram of body weight, or approximately 0.5 mg per kilogram of body weight, or approximately 1 mg per kilogram of body weight, or approximately 2 mg per kilogram of body weight, or approximately 3 mg per kilogram of body weight, or approximately 4 mg per kilogram of body weight, or approximately 5 mg per kilogram of body weight, or approximately 6 mg per kilogram of body weight, or approximately 7 mg per kilogram of body weight, or approximately 8 mg per kilogram of body weight, or approximately 9 mg per kilogram of body weight , or approximately 10 mg per kilogram of body weight.

[00339] The frequency of dosing may typically be about once per week, or about once every 2 weeks, or about once every 3 weeks.

[00340] In another embodiment, a suitable dose for administration by infusion may comprise 5-450 mg/dose, or may contain 40 mg, or 50 mg, or 60 mg per dose; or may contain 70 mg, or 80 mg, or 90 mg, or 100 mg per dose; or may contain 110 mg, or 12 0 mg, or 130 mg, or 140 mg per dose; or may contain 150 mg, or 160 mg, or 170 mg, or 180 mg per dose; or may contain 190 mg, or 200 mg, or 210 mg, or 220 mg per dose; or may contain 230 mg, or 240 mg, or 250 mg, or 260 mg per dose; or may contain 270 mg, or 280 mg, or 290 mg per dose, or may contain 300 mg, or 310 mg, or 320 mg, or 330 mg, or 340 mg, or 350 mg per dose; or may contain 360 mg, or 370 mg, or 380 mg, or 390 mg, or 400 mg, or 410 mg, or 420 mg, or 430 mg, or 440 mg, or 450 mg per dose.

[00341] In one embodiment of the invention, the dose may be delivered through one or more than one infusion. The dose may be delivered through one, two or three infusions. In some embodiments, the duration of therapy may be one or more infusions. In some embodiments, improvement in the patient's condition can be achieved through treatment over an extended period of time. In some embodiments, the duration of therapy may be until disease progression or lifelong.

[00342] In another embodiment, the pharmaceutical compositions of the present invention can be prepared in bulk form and, as such, the components of the pharmaceutical composition are present in amounts greater than may be required for administration and are suitably diluted prior to administration.

[00343] Alternatively, the pharmaceutical composition may be frozen, spray dried, or lyophilized and reconstituted in a suitable sterile carrier before use. Lyophilization can be performed using methods known in the art, which include various steps such as freezing, annealing, primary and secondary drying.

[00344] Pharmaceutical compositions can be administered as a single therapeutic agent or in combination with additional therapeutic agents as needed. Thus, in one embodiment, the present methods of treatment and/or prophylaxis are used in combination with the administration of a therapeutically effective amount of another active agent. The other active agent may be administered before, during, or after administration of the pharmaceutical compositions of the present invention. The other active agent may be administered as part of the present composition or, alternatively, as a separate formulation.

[00345] Administration of the present pharmaceutical compositions can be accomplished by a variety of routes, including parenteral, oral, buccal, nasal, rectal, and topical administration. Parenteral administration may include, without limitation, transdermal, subcutaneous, intravenous, intraarterial, intraperitoneal, intradermal, intracardiac, intraventricular, intracranial, intratracheal, intrathecal, intramuscular injection, intravitreal injection.

[00346] The pharmaceutical compositions of the present invention are particularly suitable for parenteral administration, i.e. subcutaneously, intramuscularly, intravenously, intraperitoneally, into the spinal cord, into the joints, intrasynovially and/or intrathecally. Parenteral administration can be done by bolus injection or continuous infusion. Pharmaceutical compositions for injection may be in unit dosage form, such as, but not limited to, ampoules, vials, prefilled syringes, or multi-dose containers with an added preservative. In addition, a number of recent approaches to drug delivery have been developed, and the pharmaceutical compositions of the present invention are suitable for administration by these new routes, for example, BD Physioject™, Inject-ease®, Genject®, injector pens such as GenPen®, and needleless devices such as MediJector®40 and BioJector®. The pharmaceutical composition of the present invention can also be adapted for as yet undiscovered routes of administration.

[00347] Also see Langer, 1990, Science, 249: 1527-1533.

[00348] The proposed pharmaceutical compositions can also be formulated as a depot preparation. Such long-acting formulations can be administered by implantation (eg, subcutaneously or intramuscularly) or by intramuscular injection. Thus, for example, the formulations can be modified using suitable polymeric or hydrophobic materials (eg, as an emulsion in a suitable oil), or ion exchange resins, or as sparingly soluble derivatives, eg, as a sparingly soluble salt.

[00349] Pharmaceutical compositions may optionally be provided in a vial, package, or dispenser that may contain one or more unit dosage forms containing the active ingredient. In one embodiment, the dispenser device may comprise a syringe containing a single dose of liquid formulation ready for injection. The syringe may be accompanied by instructions for administration.

[00350] In another embodiment, the present invention provides a kit or container containing an aqueous pharmaceutical composition of the invention. The concentration of the antibody in the aqueous pharmaceutical composition can vary over a wide range, but is typically within the range of about 1 to about 200 mg/ml. The kit may also come with instructions for use.

[00351] The method for obtaining the above compositions involves adding acetate buffering agents to the aqueous phase, followed by adding, in any order, the following components: trehalose, prolgolimab, and/or a solubilizer selected from the group: polysorbate 20, polysorbate 80, poloxamer 188 or their combination.

[00352] The method of obtaining the above compositions involves adding histidine buffer agents to the aqueous phase, followed by adding, in any order, the following components: trehalose, prolgolimab, and/or a solubilizer selected from the group: polysorbate 20, polysorbate 80, poloxamer 188 or their combination.

RESEARCH EXAMPLE:

[00353] The following are examples of a study to determine the reagents and their concentrations for the preparation of an aqueous pharmaceutical composition of the anti-PD-1 antibody prolgolimab of the present invention.

[00354] The studies and examples presented herein are for illustrative purposes only, which demonstrate the suitability of certain components used in aqueous pharmaceutical compositions of the anti-PD-1 antibody prolgolimab. It is understood that other methods may be used by one of ordinary skill in the art without departing from the teachings of the present invention.

[00355] The suitability of the aqueous compositions of the present invention was tested through the illustrative methods described herein.

[00356] Example 1: Preparation of stable formulations of the anti-PD-1 antibody prolgolimab

[00357] Obtaining antibody samples with a concentration of 5 mg/ml was carried out in Stirred Cell concentration cells (Millipore) under pressure. To do this, the antibody in its original composition was placed in a cell, with continuous stirring, the protein was concentrated under a stream of compressed air to a concentration of 10 mg/ml, after which at least a 10-fold volume of an aqueous solution with the target composition, including buffers, osmotic agents, and, if necessary, additional water-soluble stabilizers. Upon completion of the diafiltration process, the antibody was concentrated to a concentration of about 10 mg/ml, unloaded from the cell, and the exact protein concentration was determined by UV spectrophotometry. Then the appropriate solution of excipients was added to the sample to obtain a solution with a target protein concentration of 5 ± 0.2 mg/ml.

[00358] Protein samples with a concentration of more than 50 mg/ml were obtained in Pellicon cassettes (Millipore) in tangential flow mode. To do this, the antibody in the original composition was placed in a container for diafiltration, the protein was concentrated to a concentration of 50-100 mg/ml, after which the system was connected to a supply of at least 10 times the volume of a solution with the target composition containing buffers, osmotic agents and, if necessary, additional water-soluble stabilizers. It is allowed to add a concentrate of osmotic agents and water-soluble stabilizers after completion of diafiltration. Upon completion of the diafiltration process, the antibody was concentrated to a concentration above the target, discharged from the system, and the exact protein concentration was determined. The appropriate solution of excipients was then added to the sample to obtain a solution with the target protein concentration.

[00359] When preparing formulations containing solubilizers, surfactant concentrates were added to the antibody after completion of diafiltration and concentration with a final dilution of the antibody with a solution of excipients to the target concentration.

[00360] Before aseptic filling into the final container (eg, glass or polymer vessel, vial or syringe), the antibody solution is filtered through a 0.22 μm pore size membrane.

[00361] Example 2: Determination of protein concentration in test samples

[00362] Protein concentration was determined using UV spectrophotometry at 280 nm in UV transparent plates.

[00363] Each sample was diluted with the appropriate excipient solution to a concentration of ~0.5 mg/ml. 150 μl of the diluted sample was placed into the well of the UV spectrophotometry plate. The optical density of the solutions placed in the plate was measured using a plate spectrophotometer at a wavelength of 280 nm. The corresponding solution of excipients was used as a reference solution.

[00364] The protein concentration (C) in mg/ml was calculated using the formula:

Where

A ₂₈₀ is the optical density value at a wavelength of 280 nm;

ε is the extinction coefficient of the protein under study;

b is the total dilution factor of the sample;

l is the thickness of the layer in the well of the tablet; for 150 µl l=0.42 cm.

[00365] Example 3: PEG Aggregation Study

[00366] A solution of PEG 6000 was prepared with a mass concentration of 20-25% in the studied composition of excipients. The final solutions were filtered through a 0.4 5 µm Durapore filter.

[00367] A calculated amount of sample, excipient solution, and 20-25% PEG 6000 solution was transferred into 96-well UV spectrophotometry plates so that a number of wells had a PEG6000 concentration of 0 to 18%, and the protein concentration in each well was 1 mg/ml. All solutions obtained in the wells were mixed well by pipetting.

[00368] After this, the degree of turbidity of the solutions was assessed visually, and the optical density of the solutions was measured at a wavelength of 400 nm.

[00369] Protein precipitation in the presence of PEG is due to a volume displacement effect, that is, the protein is sterically displaced from solvent regions by the polymer. This causes the protein to concentrate until its solubility is exceeded and a precipitate forms. The less stable the sample, the lower the concentration of PEG 6000 it will form visible aggregates (opalescence).

[00370] Example 4: Determination of colloidal stability using the shake test method

[00371] The test samples were divided into 2 parts of 200 μl and placed in glass vials, 1 vial for each composition was placed in the refrigerator for storage at 2-8°C, the rest were placed in a shaker and shaken at a speed of 800 rpm at temperature 2-8°C for the specified time. After the end of the stress, the tubes were shaken on a vortex and transferred for analysis.

[00372] Example 5: Determination of colloidal stability by cryoconcentration

[00373] The test samples were divided into 2 parts and placed in polymer tubes: 1 tube for each composition was put in the refrigerator for storage at 2-8°C, the rest were placed in the freezer and stored at a temperature of minus 16-20°C for specified time. After the end of the stress, the tubes were removed from the freezer, kept at room temperature until the contents were completely thawed, the solutions were mixed using a vortex and transferred for analysis.

[00374] Example 6. Determination of thermal stability using the thermostress method

[00375] The test samples were divided into 2 parts and placed in separate glass vials: 1 vial for each composition was placed in the refrigerator for storage at 2-8°C, the rest were placed in a thermostat and incubated at the required temperature for the specified time. After heating was completed, the vials were removed from the thermostat, kept at room temperature for about 15 minutes, and submitted for analysis.

[00376] Example 7: Determination of Sample Homogeneity by Size Exclusion (SEC) High Performance Liquid Chromatography (HPLC)

Column Tosoh TSK-GelG3000SW _XL 7.8 mm ID × 30 cm, cat. No. 08541.

Column temperature: 25°C

Mobile phase flow rate: 0.7 ml/min.

Injection volume: 10 µl.

Sample concentration: 5 mg/ml.

Detector wavelength: 220 nm.

Elution duration: 25 min.

Mobile phase: Disodium hydrogen phosphate b/w 7.1 mg/ml.

Sodium chloride 17.54 mg/ml.

The pH of the mobile phase was adjusted to 7.0 with orthophosphoric acid.

The change in purity after stress was calculated using the formula:

Δ = (proportion of main peak after stress - fraction of main peak before stress)

[00377] Example 8: Determination of charge heterogeneity (charged form profile) of samples on a Caliper Labchip GX II instrument

[00378] Preparation of analyzed samples.

[00379] Samples were diluted to a concentration of 1 mg/ml. To 200 μl of the resulting solution, 2 μl of a solution of carboxypeptidase B (CpB) with a concentration of 5 mg/ml was added, mixed and incubated for 2 hours at a temperature of 37°C. The test samples were dialyzed against three changes of water. To carry out dialysis, the test solutions were placed in Amicon Ultra centrifuge tubes with a volume of 0.5 ml and centrifuged for 10 min at a rotor speed of 10,000 rpm in an Eppendorf Centrifuge 5417R centrifuge. The absorption intensity of the solutions was measured using a Cary 50 Bio spectrophotometer relative to water. Samples of the studied series were prepared with a concentration of 2 mg/ml. A 96-well Bio-Rad plate was filled with 3 µl of Labelling Buffer (from the HT Protein Charge Variant Labeling Kit), 15 µl of test solutions and 3 µl of Dye Mixture solution (from the HT Protein Charge Variant Labeling Kit). Place the plate in a dark place for 10 minutes, after which 3–6 μl of water was added to each well and mixed by pipetting. Centrifuge for 1 minute at 1000 rpm in an Eppendorf Centrifuge 5417R.

[00380] Preparation of working solutions and filling of the chip.

[00381] Preparation of working solutions and preparation of the chip were carried out in accordance with the manufacturer’s method using the HT Protein Charge Variant Labeling Kit. Running an analysis is standard procedure. Analysis method HT Protein Charge Variant 90s.

[00382] Example 9: Determination of sample purity using a Caliper Labchip GXII instrument under reducing and non-reducing conditions

[00383] Preparation of analyzed samples.

To prepare denaturing and reducing solutions, 700 μl of NT Protein Express Sample Buffer was used. To restore the samples, 24.5 μl of 1 M dithiothreitol (DTT) was added. An alkylating agent, 24.5 μl of 1 M iodoacetamide (IAM), was added to the buffer for non-reducible samples.

[00384] For each sample, two microtubes were prepared - 35 μl of denaturing buffer was added to one, and 35 μl of reducing buffer was added to the other. Samples were diluted to a concentration of 2 mg/ml. 5 μl of sample was added to each pair of tubes. Samples were denatured at 100°C for 5 minutes. The tubes were mixed by vortex, after which 70 μl of water was added to each tube and mixed by vortex. Transfer 44 μl of each sample into the wells of a 96-well plate.

[00385] Preparation of working solutions and filling the chip

Working solutions and chip preparation were carried out in accordance with the manufacturer’s method using the HT Protein Express Reagent Kit. Running an analysis is standard operation. HT analysis method Protein Express 200.

[00386] Example 10 Determination of the charged form profile of samples

ion exchange (IO) high performance liquid chromatography (HPLC) method

Column: ProPac WCX-10 Analytical, 4x250 mm.

Pre-column: Pro Race WCX-10G, 4×50 mm.

Column temperature: 30°C.

Mobile phase flow rate: 0.7 ml/min.

Injection volume: 50 µl.

Sample concentration: 1 mg/ml.

Detector wavelength: 220 nm.

Elution duration: 60 min.

Mobile phase:

Eluent A: 0.03 M 2-monfolinoethanesulfonic acid (MES), pH 6.0.

Eluent B: 0.03 M MES, 0.5 M NaCl, pH 6.0.

Eluent gradient A: 86%-0%-86%.

[00387] Before analysis, the test sample was treated with a solution of carboxypeptidase B at a temperature of +37±1°C for 2 hours.

[00388] The absolute change in the profile of charged species after stress was calculated using the formula:

Δ = | content of acidic fractions before stress - content of acidic fractions after stress | + | content of the dominant fraction before stress - content of the dominant fraction after stress | + | content of alkaline fractions before stress - content of alkaline fractions after stress.

[00389] Example 11. Determination of low molecular weight impurities by vertical gel electrophoresis (VEP) in polyacrylamide gel (PAGE) under reducing and non-reducing conditions

[00390] PAGE was prepared in glass plates in the presence of sodium dodecyl sulfate, consisting of a concentrating layer - 4% PAGE and a separating layer: under reducing conditions - 12.5% PAGE, under non-reducing conditions - 8% PAGE.

[00391] The electrophoresis chamber was assembled and installed in accordance with the operating instructions for the vertical electrophoresis device. Samples were prepared by diluting samples with purified water to a final concentration of 1 mg/mL. A volume equivalent to 40 μg was taken and the prepared samples of the test sample were mixed in a ratio of 3:1 (volume/volume) with a 4-fold sample loading buffer solution containing 2-mercaptoethanol (reducing conditions) and not containing 2-mercaptoethanol (non-reducing conditions) , mixed. The resulting solutions were incubated at a temperature of (99±1)°C for 3 minutes (samples containing 2-mercaptoethanol) and at a temperature of (99±1)°C for 1 minute (samples not containing 2-mercaptoethanol). The solutions were cooled to room temperature, mixed and applied to PAGE wells under a layer of electrode buffer solution.

[00392] Electrophoresis was carried out in constant current mode using a water cooling system. The operating parameters of the power source were set: when the dye front passed through the concentrating gel, the current voltage was 110 V. After the dye front entered the lower separating gel by 5-7 mm, the current voltage was increased to 180 V. The power source was turned off when the dye front reached the lower boundary of the gel .

[00393] After completion of electrophoresis, the gels were separated from the glasses and the proteins were fixed in a fixing solution for 16-18 hours at room temperature. Then the gels were stained (in acid blue 83 solution) and washed until the bands were clearly visualized. The gels were scanned. The purity and impurities of the test samples were assessed using GelPro software.

[00394] Example 12 Determination of relative specific activity

[00395] The relative specific activity of anti-PD-1 monoclonal antibody samples was assessed by the ability to specifically bind to the PD-1 protein on the membrane surface of Jurkat PD-1 NFAT cells. The day before the assay, PDL-1 was immobilized in the wells of culture plates. The next day, the plates were washed and 50 μl/well of phytohemagglutinin-P solution (PanEco, Russia, cat. no. M021) was added. Using the Freedom Evo robotic station, serial dilutions of the standard and test samples were prepared and added to culture plates at 10 μl/well. 40 μl/well of Jurkat PD-1 NFAT cell suspension was added to culture plates. The plates were incubated for 4-6 hours at 37°C, 5% CO ₂ . All procedures described above were carried out under aseptic conditions.

[00396] Upon completion of incubation, 100 μl/well of BioGlo solution (Promega, USA, cat. no. G7941) was added and the luminescence level was determined.

[00397] Using Magellan software, four-parameter curves of the dependence of the average luminescence value on the protein concentration were constructed for solutions of the standard sample and the test samples located on the same plate.

[00398] The relative specific activity of the test sample in percent (RP) was calculated using the formula: , Where

ED ₅₀ st - the value of the half effective dose of the standard sample, ng/ml;

ED ₅₀ test - the value of the half effective dose of the test sample, ng/ml.

[00399] The final result was taken as the average value of the relative specific activity, calculated from three independent determinations (determined on 3 different culture plates).

[00400] Example 13 Viscosity Determination

The dynamic viscosity of the solutions under study was determined by rotational viscometry on a Brookfield CAP2000+L viscometer.

[00401] Example 14: Investigation into the nature of the buffer solution

[00402] Test compounds

Four buffer solutions were selected for this study. When choosing the molarity and pH of the buffer, restrictions on possible subcutaneous administration were taken into account, as well as the pI of the antibody (the pH of the buffer solutions under study was selected at the minimum possible physiological value).

Composition (per 1 ml):

[00403] Determination of colloidal stability by PEG aggregation method.

The study was carried out in triplicate for each image. The results are presented in Table 1 and Fig. 1.

[00405] As a result of the study, samples in histidine and acetate buffer solutions showed the highest colloidal stability in the presence of PEG. Citrate and phosphate formulations were excluded from further study because aggregation at 6% PEG is an indication of poor colloidal stability. Based on the results obtained, acetate and histidine buffer solutions were selected for further selection of pH and molarity of the buffer solution.

[00406] Example 15. Selection of pH and buffer capacity of the solution

[00407] Test formulations (per 1 ml)

[00408] The study was carried out using the thermostress method at 50°C for 72 hours. The results are presented in Table 2. The homogeneity of the samples was analyzed by E-HPLC and electrophoresis on a Labchip device. The profile of the charged forms of the samples was analyzed on a Labchip device.

[00410] The study showed that the presence of a monoclonal antibody against PD-1 (prolgolimab) in an aqueous solution leads to an increase in pH levels. The greatest pH stabilization is provided by 20 mM buffer solutions.

[00411] All studied samples demonstrated high aggregation stability on E-HPLC, since the increase in impurities during thermal stress was no more than 0.5% for all samples. The best stability was shown by samples based on histidine with pH 5.5-6.0, as well as on acetate with pH 5.0-5.5.

[00412] All formulations demonstrated similar stability of the profile of charged forms, the quantitative difference in the change in fraction content between formulations was within the error limits of the method.

[00413] According to the results of gel electrophoresis under reducing conditions, all histidine-based formulations demonstrated higher stability, while under non-reducing conditions, the best performance was obtained for acetate buffer solutions.

[00414] Based on the obtained stability data in terms of pH, purity and profile of the charged forms of the monoclonal antibody against PD-1, the following compositions of excipients are recommended for further work:

[00415] Example 16: Selection of Histidine-Based Pharmaceutical Composition

[00416] Test compounds

To screen a stable pharmaceutical composition based on a 20 mM histidine buffer solution with pH 5.5, the following excipients were taken: mannitol, trehalose dihydrate, sucrose (osmotic agents). All tested solutions are isotonic.

[00417] Study formulations (mg to 1 ml)

[00418]

The stability of the samples was studied using the thermostress method at 50°C for 72 hours, the shake test for 120 hours at a speed of 800 rpm, as well as single freezing at a temperature of minus 16-20°C and thawing at a temperature of +25±1° WITH. The turbidity of solutions was assessed by spectrophotometry at 400 nm. The homogeneity of the samples was analyzed by E-HPLC and electrophoresis on a Labchip device. The profile of the charged forms of the samples was analyzed on a Labchip instrument.

[00421] Table 3 shows the negative effect of mannitol on

thermal and colloidal stability of the monoclonal antibody against PD-1 in a 20 mM histidine buffer solution with pH 5.5: during thermal stress for 96 hours, the increase in impurities on E HPLC ranged from 1.24 to 3.18% during a shake test for 120 hours, the solutions became visible aggregation. Compositions with mannitol also showed negative stability results during cryoconcentration: after one freeze-thaw cycle, the increase in impurities on E-HPLC ranged from 1.05 to 4.45%, which is much higher than this figure for other compositions.

[00422] Formulations containing L-proline also demonstrated poor colloidal stability due to the formation of visible aggregation during the shake test. After freezing-thawing and thermal stress tests, the increase in impurities of formulations with L-proline averaged more than 1% on E-HPLC.

[00423] The experiment revealed high thermal and colloidal stability of prolgolimab (no significant changes in quality indicators under all stress influences) in formulations with trehalose dihydrate without the presence of solubilizers, such as polysorbate 80 and poloxamer 188.

[00424] The greatest stabilizing effect on the monoclonal antibody against PD-1 in a 20 mM histidine buffer solution is exerted by trehalose dihydrate, sucrose, and their combinations with glycine. These formulations can be used for a lyophilic dosage form of a monoclonal antibody.

[00425] Evaluation of the stability of promising histidine-based formulations during freeze-drying

[00426] The samples that demonstrated the best stability in the previous screening stage were examined for the possibility of obtaining a lyophilisate for the preparation of a solution for infusion.

[00427] For this purpose, solutions containing 20 mg/ml or 100 mg/ml of the monoclonal antibody to PD-1 prolgolimab were poured into hydrolytic class I glass vials, the vials were loosely sealed with a rubber stopper with a slot. Vials with the solution were placed in a freeze-drying chamber. Freeze drying was carried out in automatic mode. The freezing stage was carried out at a temperature of minus 40°C, primary drying was carried out at a pressure of (0.10±0.03) mbar, during secondary drying the temperature was increased, the pressure was (0.05±0.02) mbar. After completing the drying process, the pressure was reduced to the required vacuum value of 0.76 ± 0.03 mbar, the bottles were sealed with rubber stoppers and then the vacuum was released to atmospheric pressure. The sealed vials were transferred for further research. Lyophilized samples of the anti-PD-1 monoclonal antibody prolgolimab were stored at 2-8°C.

[00428] To confirm protein stability after lyophilization, samples were reconstituted. The monomer content was assessed by E-HPLC, and the profile of charged forms was analyzed on a Labchip instrument. The pH value was also determined before and after lyophilization. The results are presented in Table 5.

[00429] When freeze-drying selected histidine-based formulations and after reconstituting the lyophilisates, a slight pH shift was observed, within the error limits of the method. All formulations demonstrated high stability in terms of E-HPLC purity and charged species profile after reduction. However, a noticeable difference in the purity of the samples was demonstrated by gel electrophoresis in the LabChip Caliper system. The most stable samples in terms of this indicator are compositions No. 22, 27, 36, containing trehalose dihydrate. Formulations in L-proline also showed a significant change in the profile of charged forms and are therefore not recommended for use. The possibility of lyophilization of formulations No. 22 and 27 with a protein concentration of 100 mg/ml was demonstrated.

[00431] Obtaining a highly concentrated form and confirming stability during accelerated storage

[00432] Based on the results of screening in liquid and lyophilic dosage forms, the following formulations were selected for accelerated stability studies:

[00433] The trehalose dihydrate content was reduced to equalize the osmolality of solutions with increased concentrations of anti-PD-1 monoclonal antibody to physiological levels (about 300 mOsm/kg), as well as to reduce the viscosity of solutions to less than 100 cP. The stability of the samples was confirmed by accelerated storage at +37°C, the analysis was carried out by E HPLC, IO HPLC and VEF, and the relative specific activity of prolgolimab was determined. The results are presented in Tables 6 and 7.

[00436] Example 17: Selection of an acetate-based pharmaceutical composition

[00437] Test compounds

To screen a stable pharmaceutical composition based on a 20 mM acetate buffer solution with pH 5.0, the following excipients were taken: mannitol, trehalose dihydrate, sucrose (osmotic agents), L-proline (osmotic agent and stabilizer), glycine (osmotic agent and stabilizer), polysorbate 80 and poloxamer P18 8 (solubilizers). All tested solutions are isotonic.

[00438] Test formulations (per 1 ml).

[00439] The stability of the samples was studied using the thermostress method at 50°C for 72 hours, the shake test for 120 hours at a speed of 800 rpm, as well as single freezing minus 16 - 20°C and thawing at a temperature of +25±1 °C. The turbidity of solutions was assessed by spectrophotometry at 400 nm. The homogeneity of the samples was analyzed by E-HPLC and electrophoresis on a Labchip device. The profile of the charged forms of the samples was analyzed on a Labchip instrument.

[00442] As a result of the work, the negative effect of mannitol on the thermal and colloidal stability of the monoclonal antibody against PD-1 in a 20 mM acetate buffer solution with pH 5.0 was shown: during thermal stress for 96 hours, the increase in impurities on E HPLC ranged from 0.48 to 2.59%, During the shake test for 120 hours, all the solutions under study did not acquire visible aggregation (see the results of UV spectrophotometry). Compositions with mannitol also showed negative stability results during cryoconcentration: after one freeze-thaw cycle, the increase in impurities on E-HPLC ranged from 0.69 to 9.44%, which is much higher than this figure for other compositions.

[00443] Formulations containing L-proline also demonstrated low thermal stability in terms of purity when studied by E-HPLC: the increase in impurities after 96 hours of stress was 1.64-1.93%. After freezing-thawing, the increase in impurities in formulations with L-proline did not exceed the average value of the most stable formulations in the group. The experiment did not reveal the effect of solubilizers on the thermal or colloidal stability of the protein. Prolgolimab in formulations with trehalose dihydrate showed high stability under all stress conditions in the absence of surfactants such as polysorbate 80 and poloxamer 188.

[00444] The greatest stabilizing effect on the monoclonal antibody against PD-1 in a 20 mM acetate buffer solution has: trehalose dihydrate, sucrose, and their combinations with glycine. These compositions can be used to prepare a lyophilic dosage form of a monoclonal antibody.

[00445] Determination of the stability of promising acetate-based formulations during freeze drying

[00446] The samples that demonstrated the best stability in the previous screening stage were examined for the possibility of obtaining a lyophilisate for the preparation of a solution for infusion.

[00447] To do this, solutions containing 20 mg/ml or 100 mg/ml of monoclonal antibody against PD-1 were poured into glass vials of hydrolytic class I, the vials were loosely sealed with a rubber stopper with a slot. Vials with the solution were placed in a freeze-drying chamber. Freeze drying was carried out in automatic mode. The freezing stage was carried out at a temperature of minus 40°C, primary drying was carried out at a pressure of (0.10±0.03) mbar, during secondary drying the temperature was increased, the pressure was (0.05±0.02) mbar. After completing the drying process, the pressure was reduced to the required vacuum value of 0.76 ± 0.03 mbar, the bottles were sealed with rubber stoppers and then the vacuum was released to atmospheric pressure. The sealed vials were transferred for further research. Lyophilized anti-PD-1 monoclonal antibody samples were stored at 2-8°C.

[00448] To confirm protein stability after lyophilization, samples were reconstituted. The monomer content was assessed by E-HPLC, and the profile of charged forms was analyzed on a Labchip instrument. The pH value was also determined before and after lyophilization. The results are presented in Table 8.

[00449] When freeze-driing selected acetate-based formulations and after reconstituting the lyophilisates, a significant pH shift in the range of 0.26-0.45 is observed. This shift may be due to the loss of acetate ions during the drying process. Thus, acetate-based compositions can be recommended for freeze-drying, provided that this phenomenon is taken into account.

[00450] All formulations demonstrated high stability in E-HPLC purity and charged species profile after reduction. During the work, the possibility of lyophilization of compositions No. 10 and 12 with a protein concentration of 100 mg/ml was demonstrated.

[00452] Obtaining a highly concentrated form and confirming stability during accelerated storage

[00453] Based on the results of screening in liquid and lyophilic dosage forms, the following formulations were selected for accelerated stability studies:

[00454] The trehalose dihydrate content was reduced to equalize the osmolality of solutions with increased concentrations of anti-PD-1 monoclonal antibody to physiological levels (about 300 mOsm/kg), as well as to reduce the viscosity of solutions to less than 100 cP. The samples were stored at +37°C and analyzed by E HPLC, IO HPLC and VEF methods, and the specific activity of the protein was determined. The results are presented in Tables 9 and 10.

[00457] While the invention has been described with reference to preferred embodiments, it should be understood that changes and modifications may be made as will be apparent to those skilled in the art. Such changes and modifications should be considered within the scope and scope of the present invention.

[00458] Illustrative, non-limiting examples of implementation of the present invention have been described in detail above. This detailed description is provided merely to provide one skilled in the art with additional details for the practice of preferred aspects of the present teachings, and is not intended to limit the scope of the present invention. In addition, each of the additional features and ideas disclosed above and below may be used individually or in combination with other features or ideas.

[00459] Moreover, combinations of features and steps disclosed in the following detailed description, as well as experimental examples, may not be necessary for the practice of the invention in the broadest sense, and are provided only to describe in detail specific examples for the present inventions. Moreover, the features of the specific examples described above and the independent and dependent claims set forth herein may be combined in ways not specifically and directly indicated to realize additional useful embodiments of the present invention.

[00460] Additional examples of studies have been conducted using various aqueous pharmaceutical compositions containing the anti-PD-1 antibody prolgolimab for the treatment of malignancies such as melanoma, including unresectable or metastatic melanoma, early stages of melanoma before and after radical treatment, cancer lung, non-small cell lung cancer (NSCLC), including unresectable or metastatic non-small cell lung cancer. Typical pharmaceutical compositions used for these studies are characterized by the following compositions below (Table 12.1).

[00461] Basic information about the clinical development of the investigational drug

The pharmacokinetics, pharmacodynamics, safety and immunogenicity of BCD-100 (prolgolimab) as monotherapy when administered intravenously were studied in a phase I dose-escalation clinical trial. This study demonstrated the safety and benefits of the study product in patients with common forms of malignant neoplasms of various localizations (melanoma, NSCLC) and, based on the results of the study, two modes of administration of BCD-100 were selected for further clinical development, 3 mg/kg IV 1 time at 3 weeks and 1 mg/kg IV once every 2 weeks.

[00462] The objective of Study No. BCD-100-2/MIRACULUM of BCD-100 in patients with unresectable or metastatic melanoma was to evaluate the pharmacokinetics, efficacy, safety and immunogenicity of monotherapy in two administration regimens: 3 mg/kg IV once a day. 3 weeks and 1 mg/kg IV once every 2 weeks. The primary endpoint (ORR) was assessed at the interim analysis at 6 months. BCD-100 demonstrated a favorable safety profile across all dose regimens. Both dose regimens demonstrated efficacy of approximately 60% CR and 30% ORR in patients with unresectable or metastatic melanoma.

[00463] Example 18. Use of prolgolimab (BCD-100) for the treatment of patients with advanced forms of malignant neoplasms of various sites, phase I study, BCD-100-1

[00464] Study Design

Study No. BCD-100-1 examined the pharmacokinetics and tolerability of BCD-100 (prolgolimab). Study No. BCD-100-1 (NCT03050047) was a multicenter, open-label, phase I study in patients with solid tumors conducted in the Russian Federation. The primary objective of the study was to evaluate the pharmacokinetics and clinical pharmacological parameters of BCD-100. The main characteristics of the BCD-100-1 study are shown in Table 13.

[00466] The first patient was prescribed a starting dose of BCD-100 of 0.3 mg/kg every two weeks. In the absence of dose-limiting toxicity within 4 weeks, the patient's dose was increased to 1 mg/kg every two weeks. After the first patient, the study continued according to the classical design with dose escalation 3+3, i.e. in the absence of EBRT, a cohort of 3 patients was sequentially enrolled into the study every 4 weeks at the next dose level. At each dose level, BCD-100 was administered for 85 days (about 3 months) or until signs of DLT or disease progression appeared.

[00467] If DLT events at the current dose level were observed in one patient, the next cohort of three patients received the drug at the same level, that is, a total of six patients began receiving a given dose of BCD-100. If DLT events were observed in two or more patients at the same dose level, dose titration and administration of the drug to new patients were stopped.

[00468] The duration of follow-up for patients after therapy with BCD-100 was 126 days. During this period, monitoring of adverse events, medical examinations, and blood tests were carried out (once every 2 weeks - the first 28 days, then once every 28 days).

[00469] Study results

Summary of Patient Population Characteristics

The study included 15 patients with common forms of malignant solid neoplasms of various localizations (melanoma, including choroidal melanoma, NSCLC, renal cell cancer, mesothelioma), aged 18 years and older, of both sexes. To be included in the study, patients had to have an ECOG score of 0-2 and at least one measurable target lesion according to RECIST 1.1 (not including bone metastases). A description of the disease characteristics of the patients and the distribution of patients is presented in Table 14, Table 15.

[00472] The final analysis included data from 15 patients (6 patients from the BCD-100 1 mg/kg cohort, including 1 patient after dose escalation, 6 patients from the BCD-100 3 mg/kg cohort, and 3 patients from the BCD-100 cohort 10 mg/kg). All patients received therapy during the main phase of the study (85 days).

[00473] All patient cohorts were balanced for basic demographic characteristics. The distribution of all patients by gender was even: women - 53.33% (8 patients), men - 46.67% (7 patients). The median age of patients was 56 years (minimum 35 years, maximum 77 years).

[00474] According to the characteristics of the underlying disease, patients with melanoma predominated in the population (9/15 patients). The population also included 4 patients with NSCLC, 1 patient with pleural mesothelioma, and 1 patient with renal cell carcinoma. The duration of the disease at the time of screening in the entire population was (median) 17.07 months, minimum - 0.6 months, maximum - 91.2 months.

[00475] Summary of clinical safety information from the Phase I study

Safety was examined in a population that included all patients who received at least one dose of study drug (n=15). Each of the 15 patients had AEs and/or SAEs, the total number of events was 247. Statistical analysis did not reveal differences between the cohorts in the number of AEs (p = 0.567, Kruskal-Wallis test). Summary Table 16 with safety data for the drug is shown below.

[00476] AEs corresponding to grade 3 severity according to the CTCAE 4.03 scale were noted in 40.00% (6 of 15) of patients - two patients in each cohort. In 4 of 6 of these patients, AEs grade 3 or higher were considered treatment related by the investigator: in a patient receiving BCD-100 at a dosage of 1 mg/kg, in a patient receiving BCD-100 at a dosage of 3 mg/kg, and in two patients receiving BCD-100 at a dosage of 10 mg/kg. Many of these adverse events were hematologic in nature.

[00477] SAEs were reported in three patients receiving 1 mg/kg BCD-100 and one patient receiving 3 mg/kg BCD-100.

[00478] One case of dose-limiting toxicity (DLT) was identified during the study. Patient 13-09, who received two doses of BCD-100 at a dosage of 3 mg/kg, had hormonal imbalance (TSH decreased) and developed autoimmune thyroiditis (immune-mediated adverse event of grade 2 according to CTCAE 4.03 criteria). The Data and Safety Monitoring Board considered this case to be a DLT event. Patient 13-0 9 continued to receive study therapy, and the course of his disease was characterized as stable according to RECIST 1.1 and irRC criteria.

[00479] In addition to autoimmune thyroiditis, immune-mediated AEs were noted in 33.33% (5 of 15) of patients: 33.33% (2 of b) of patients receiving 1 mg/kg BCD-100, 50.00% ( 3 of 6) patients receiving 3 mg/kg BCD-100, but none of the patients receiving 10 mg/kg BCD-100. All of these AEs were grade 1 events according to the CTCAE 4.03 scale, and were not considered to be DLT events.

[00480] AEs/SAEs resulted in temporary discontinuation of study therapy in three patients: 16.67% (1 of 6) of patients receiving 1 mg/kg BCD-100 and 33.33% (2 of 6) of patients receiving 3 mg/kg BCD-100. All AEs due to which therapy was temporarily discontinued were of an immune nature (grade 1 hyperthyroidism in patient 10-03, who received 1 mg/kg BCD-100; grade 1 autoimmune thyroiditis in patient 13-10, and grade 2 autoimmune thyroiditis in patient 13-09 - both patients received 3 mg/kg BCD-100). Patient 10-03's investigational therapy was suspended. Patients 13-09 and 13-10 were prescribed glucocorticoids for the disease, after which study therapy was resumed. None of the breaks in therapy exceeded the norms (2-4 weeks) established in the study protocol.

[00481] In three patients, the study drug was discontinued due to disease progression (two of them after 5 administrations at a dosage of 1 mg/kg and one after 5 administrations at a dosage of 10 mg/kg). The reason for discontinuation in all cases was disease progression and not BCD-100 use.

[00482] The SAE resulted in discontinuation of therapy and subsequently the death of one of the subjects. Patient 13-10, who underwent three injections of the drug at a dosage of 3 mg/kg, experienced a grade 5 hemorrhage in the right hemisphere. The investigator considered that this AE was likely related to the study treatment.

[00483] There were no differences between the patient groups in terms of changes in laboratory and physiological parameters over time. Although analysis of these parameters revealed isolated examples of statistically significant differences, these differences were considered to reflect natural variability within the normal range. Abnormalities detected by the results of a biochemical blood test and a blood coagulation test (increased levels of transaminases, bilirubin, abnormalities in blood clotting parameters, etc.) in most cases were typical for the study population (patients with inoperable neoplasms), expected, and transient ; normalization of parameters passed without consequences; no additional therapy was required.

[00484] The described results demonstrate a favorable safety profile of BCD-100 at any of the studied dosages when used intravenously.

[00486] Immunogenicity

Immunogenicity assessments were performed at screening, on day 28 of study therapy, at the end of the study period (on day 85), and among patients continuing to receive BCD-100 therapy, also every 42 days thereafter. Immunogenicity assessment was carried out in two stages: the first stage was screening for the content of drug binding antibodies (CAT) in patient serum samples, and the second stage was screening of samples with a positive CAT titer for the content of neutralizing antibodies (HAT).

[00487] All patients whose serum samples were available for analysis at the time of screening and on subsequent testing days were included in the immunogenicity analysis (n=15). No samples were found to contain CAT. Due to the lack of samples with CAT, HAT analysis was not performed.

[00488] Summary of pharmacokinetics and metabolism information obtained during the phase I study

The pharmacokinetics (PK) analysis included data from patients who did not have more than 3 blood serum samples missed/lost/spoilt to determine the concentration of BCD-100 after the first administration of the drug (n=15).

[00489] Pharmacokinetics studies (both after the first administration of the drug and throughout all administrations) were carried out in the following groups:

• in the first patient (BCD-100 dose 0.3 mg/kg) (n=1);

• in the first cohort of patients (BCD-100 dose 1 mg/kg) (n=5);

• in the second cohort of patients (BCD-100 dose 3 mg/kg) (n=6).

• in the third cohort of patients (BCD-100 dose 10 mg/kg) (n=3).

[00490] The pharmacokinetic analysis included evaluation of standard PK characteristics of the distribution and elimination of the study drug after repeated intravenous administration. The results of the analysis are presented in Table 17.

[00491] Blood samples for pharmacokinetics analysis (serum BCD-100 levels) were collected from all patients enrolled in the study at each dose level. The following time points for sampling were used: before the first administration of the drug; after 30 minutes, 2 hours (±15 minutes), 4 hours (±15 minutes), 6 hours (±15 minutes), 24 hours (±1 hour), 48 hours (±2 hours), 192 hours (±8 hours ), and 336 hours (±8 hours) after the first administration of the drug (before the second administration), and before each subsequent administration of the drug once every two weeks. Blood samples were collected for analysis of serum BCD-100 concentrations at the same time as samples were taken for immunogenicity analysis (to assess the possible effect of antibodies to the drug on the pharmacokinetic parameters of BCD-100). The concentration of BCD-100 in blood serum was determined using a validated ELISA method.

[00493] All parameters were measured after a single administration of the drug, with the exception of the C _min indicator, which was calculated for 6 administrations of the drug (12 weeks).

[00494] Statistically significant differences between the three dose cohorts were noted for AUC _{(0-336 hours)} (p=0.0088, Kruskal-Wallis test); AUMC _{(0-336 h)} (p=0.0088, Kruskal-Wallis test); AUC _(o-∞) (p=0.0123, Kruskal-Wallis test); Cmax (p=0.0103, Kruskal-Wallis test). More favorable pharmacokinetic profiles were observed at doses of 3 mg/kg and 10 mg/kg compared to the 1 mg/kg dose; however, PK, efficacy and safety indicators were not associated with dosage.

[00495] BCD-100 drug concentration profiles from the first dose to week 10 in all dose cohorts (1 mg/kg cohort [n=5], 3 mg/kg cohort [n=6], and 10 mg/kg cohort kg [n=3]) are shown in figure 1 and figure 2.

[00496] Calculation of PK parameters showed that the concentration of BCD-100 varies directly proportional to the administered dose, reaching a peak between 30 minutes and 6 hours after administration and gradually decreasing thereafter. There was no relationship between the half-life and the amount of drug administered. Indicator value turned out to be typical for monoclonal antibodies (12-18 days).

[00497] Summary of pharmacodynamic information obtained during the Phase I study

All patients whose serum samples were available for analysis at the time of screening and on subsequent testing days were included in the pharmacodynamic analysis (n=15).

[00498] The pharmacodynamics study was conducted in the following groups:

• in the first patient (BCD-100 dose 0.3 mg/kg) (n=1);

• in the first cohort of patients (BCD-100 dose 1 mg/kg) (n=5);

• in the second cohort of patients (BCD-100 dose 3 mg/kg) (n=6).

• in the third cohort of patients (BCD-100 dose 10 mg/kg) (n=3).

[00499] BCD-100 PD-1 receptor saturation measures the drug's interaction with its target and is the most important pharmacodynamic marker of anti-PD-1 monoclonal antibody activity because blockade of the PD-1 receptor initiates an antitumor immune response. Blood sampling to assess pharmacodynamics (% saturation of the PD-1 receptor with BCD-100) was performed in all patients. Samples for PD assessment were taken before drug administration, 4 hours and 336 hours after administration (but before the second administration), and before the sixth drug administration.

[00500] The results demonstrate high saturation of the PD-1 receptor with BCD-100 (95% to 100%) at all dose levels and in all cell populations (Table 18).

[00502] Summary of Clinical Efficacy Information from Phase I Study

Fourteen of 15 patients receiving BCD-100 were included in the efficacy population. One patient had treatment discontinued due to a SAE (death); At the time of the patient's death, tumor response to therapy had not been assessed using CT.

[00503] Overall response rate (partial response rate + complete response rate) and disease control rate (stable disease + partial response + complete response) were determined after 85 days of BCD-100 therapy. The efficacy assessment after 85 days of treatment with BCD-I00 was based on analysis of CT scan results. Tumor response was assessed using RECIST 1.1 and Immune-Related RECIST (irRECIST) criteria.

[00504] According to the irRECIST criteria, which were developed with immunotherapy in mind, disease control was achieved in 28.57% (4 of 14) of patients. The overall response rate was 7.14% (1 of 14). There were no significant differences in response rates between the different patient cohorts (Table 19).

[00506] Output

The analysis of the data obtained clearly demonstrates the favorable safety profile of the drug BCD-100 when used in a wide range of doses from 0.03 mg/kg to 10.0 mg/kg. For further clinical study, it seems optimal to choose 2 administration regimens for BCD-100: 1 mg/kg once every 2 weeks and 3 mg/kg once every 3 weeks.

[00507] Example 19: Prolgolimab (BCD-100) for the Treatment of Patients with Unresectable/Metastatic Melanoma, Phase II Study, BCD-100-2/MIRACULUM

[00508] Study Design

Study No. BCD-100-2/MIRACULUM (MIRACULUM, NCT03269565) is a multicenter, open-label, randomized study of the pharmacokinetics, efficacy, safety and immunogenicity of BCD-100 (prolgolimab) as monotherapy in patients with unresectable/metastatic melanoma, previously untreated or treated. . Comparisons were made between BCD-100 administered intravenously at a dose of 3 mg/kg every three weeks and BCD-100 administered intravenously at a dose of 1 mg/kg every two weeks. The study is currently ongoing in the Russian Federation and Belarus. Data for 1 year of therapy were obtained and analyzed.

[00509] Each performance parameter was assessed separately for each group. The primary efficacy endpoint was overall response rate (partial response + complete response rate) by irRECIST in study participants receiving BCD-100. The best response to therapy over the above period was taken into account when calculating ORR (overall response rate) and disease control. Secondary endpoints for assessing the effectiveness were progression-free and overall survival of patients 12 months after the start of therapy, the rate of achieving disease control (rate of achieving stabilization + partial response + complete response), time to achieve a response to therapy, duration of response to therapy (Figure 3, table 20).

[00511] Study results

Summary of Patient Population Characteristics

[00512] The study included 131 patients over the age of 18 years. The patient population included both men and women with advanced melanoma, including choroidal melanoma. This population included patients who took the place of those who dropped out before the first administration of BCD-100 or due to serious deviations from the Protocol. The resulting modified ITT population, consisting of patients who received at least one dose of BCD-100, consisted of 126 patients. To be included in the study, patients had to meet an EC0G score of 0-1 and have at least one measurable target lesion according to RECIST 1.1 (excluding bone metastases) (Figure 4, Table 21).

[00514] The safety analysis included all patients who received at least one administration of study drug (modified intent-to-treat (mITT) population, n=126).

[00515] Efficacy analysis was conducted in two patient populations:

• all patients who received at least one administration of the study drug (mITT population, n=126);

• patients who received at least one injection of the study drug and underwent at least the first scheduled CT study over time to assess response (reg protocol (PP) population, n=114).

[00516] The pharmacokinetic analysis included data from patients who received at least one administration of BCD-100 and did not have missing/lost/spoilt required samples according to the current version of the protocol at the time of determination (n=125).

[00517] Summary of Clinical Effectiveness Information from Study No. BCD-100-2/MIRACULUM

The results of the primary endpoint efficacy assessment (ORR) indicate sufficient effectiveness of BCD-100 in both dose regimens in all patient populations studied. Thus, ORR and disease control in the RR population were 40.68% and 67.80% in group 1 and 32.73% and 52.73% in group 2 of therapy. Similar efficacy rates were observed in the mITT population. Thus, the expected target response rate (28%) and the critical r value (11 responses) were achieved in both treatment groups.

[00518] A subgroup analysis by treatment line demonstrated high efficacy of BCD-100 at a minimum dose of 1 mg/kg in a population of previously untreated patients (ORR was 50.00%).

[00519] The results of the efficacy analysis for secondary endpoints confirmed the conclusion that BCD-100 was sufficiently effective at both doses. The DFS, OS, and duration of response rates were comparable to those of the best-in-class PD-1 inhibitors.

[00520] The 12-month DFS was 41.27% for patients in the BCD-100 1 mg/kg group and 34.92% for patients in the BCD-100 3 mg/kg group. Median PFS was 5.78 months (95% CI 3.52 - -) for patients in the BCD-100, 1 mg/kg group and 2.33 months (95% CI 2.07-10.25) for patients in the BCD group -10C, 3 mg/kg (p=0.4 00, log-rank test). A detailed assessment of progression-free survival in each group, according to lines of therapy (according to irRECIST criteria, in the mITT population), did not reveal statistically significant differences. BCD-100 was equally effective when used in treatment-naive patients and when used in previously treated patients.

[00521] At a median follow-up of 13.8 months (95% CI 13.2-14.7), the median overall survival for patients in the BCD-100 group, 1 mg/kg, was not reached (95% CI - ¹ Here and beyond) in the text "-" means "not achieved" - -). The 12-month OS was 74.60% for patients in the BCD-100 1 mg/kg group. The median OS for patients in the BCD-100, 3 mg/kg group was 15 months (95% CI 9.99 - -) with a median follow-up of 14.5 months (95% CI 13.9-15.2). The 12-month OS was 53.97% for patients in the BCD-100 3 mg/kg group. A detailed assessment of overall survival in each group, according to lines of therapy, did not reveal statistically significant differences. BCD-100 was equally effective when used in treatment-naive patients and when used in previously treated patients. Median OS was not achieved in any of the subgroups in patients receiving BCD-100 at a dose of 1 mg/kg (p=0.800; log-rank test). Median OS was 16.8 months (95% CI 9.33 - -) for previously untreated patients and 15 months (95% CI 7.4 6 - -) for previously treated patients in the BCD-100 3 mg group /kg (p=0.900; log-rank test) (table 22, table 23, table 24, table 25, figure 5, figure 6, figure 7, figure 8).

[00523] Summary of Clinical Safety Information from Study No. BCD-100-2/MIRACULUM

BCD-100 demonstrated a favorable safety profile at both dose regimens (Table 23).

[00525] Based on the above data, it can be concluded that the results obtained in the present study do not contradict the known data on the safety profile of monoclonal antibodies to the PD-1 receptor. There were no statistically significant differences in the safety profile of the drug at the studied doses, with the exception of the number of SAEs followed by death (a significantly higher number was noted among patients in the BCD-100 group, 3 mg/kg).

[00526] Immunogenicity

All patients whose serum samples were available for analysis at the time of screening and on subsequent testing days were included in the immunogenicity analysis (n=121). None of the patients had CAT to BCD-100.

[00527] Summary of pharmacokinetics and metabolism information from Study No. BCD-100-2/MIRACULUM

The analysis showed that a single intravenous administration of the drug BCD-100 at a dose of 1 to 3 mg/kg is characterized by a linear increase in the concentration of the test substance in the blood plasma, followed by a uniform decrease, while the half-life of the drug, regardless of the dose administered to the body, is characterized by The usual duration for immunoglobulins of the IgG class is from 11.5 to 17 days (Figure 9, Figure 10).

[00528] The trend towards a dose-dependent increase in the concentration of the test substance in the blood plasma (according to the parameters Cmax and AUC _0-t,ss ) also persists after repeated administration of the drug BCD-100, which corresponds to the literature data on the PK profile of other anti-PD-1 drugs antibodies.

[00529] The persistent preservation of C _min at a sufficient level during the course of treatment demonstrated in clinical study No. BCD-100-2/MIRACULUM indicates that therapeutic concentrations of the drug are maintained as when using the drug BCD-100 at a dose of 1 mg/kg 1 time every 2 weeks, and when using the drug at a dose of 3 mg/kg in a regimen of 1 time every 3 weeks. Thus, the use of BCD-100 in both dose regimens is justified from the standpoint of pharmacokinetic properties.

[00530] The assessment of the T1 _/2 parameter, including a comparative assessment between treatment groups, was difficult due to the lack of a typical terminal period of the drug elimination phase in a number of patients after a single administration.

[00531] In general, BCD-100, both at a dose of 1 mg/kg every 2 weeks and at a dose of 3 mg/kg every 3 weeks, was characterized by pharmacokinetic parameters that ensured a long stay of the drug in the body sufficient to maintain a stable therapeutic concentrations against the background of repeated administration. No dose dependence was observed regarding the efficacy and safety of the drug.

[00532] Summary of pharmacodynamics information obtained during the Phase I study

[00533] The percentage of PD-1 receptor saturation by BCD-100 in all leukocyte subpopulations studied did not differ between treatment groups. PD-1 saturation of >99% in the population of activated T helper cells and cytotoxic leukocytes was recorded in 33.33% of patients included in the analysis of this indicator (14 of 42). However, there was no statistically significant difference between the treatment groups: 8 patients received BCD-100 at a dose of 1 mg/kg, 6 patients at a dose of 3 mg/kg.

[00534] Ki-67 is a universal marker of cell proliferation, which is due to its presence in the cell only during the cell division process and destruction within 1.5-2 hours after the end of mitosis. The Ki-67 marker is detected in the telomere region, centromeres and nucleoli. The percentage of Ki-67 positive cytotoxic T lymphocytes increased during therapy in both groups, but no significant differences were found. Analysis of the proportion of Ki-67 positive cytotoxic T lymphocytes also did not reveal statistically significant differences between the groups.

[00535] The percentage of Th9 in the total population of T helper cells during therapy increased mainly in group 2 (BCD-100, 3 mg/kg Q3W), but no significant differences were found. There were no significant differences in intergroup comparisons.

[00536] According to the available literature, early increase in Th9 levels is associated with a better response to nivolumab therapy in melanoma ³ ( ). It should be noted that these results were obtained on a limited sample of patients (n=42), heterogeneous in gender and characteristics of the underlying disease.

[00537] A comparative analysis of the proportion of Th9 in the total population of T helper cells in subgroups with different types of response to therapy with BCD-100 did not reveal statistically significant differences in both dose cohorts. However, the presented graphs show a tendency towards a better response to therapy in patients with initially elevated Tn9 levels (Figure 11, Figure 12).

[00538] Output

[00539] The data obtained on the effectiveness, safety and pharmacokinetic properties of BCD-100 are sufficient to justify the use of the drug both in a dose regimen of 1 mg/kg once every 2 weeks and in a dose regimen of 3 mg/kg once every 3 weeks.

[00540] The international, multicenter, randomized, open-label, phase II study No. BCD-100-2/MIRACULUM in a population of patients with unresectable and metastatic melanoma demonstrated significant therapeutic benefits compared to known chemotherapy efficacy data, and results comparable to the best existing treatments . Taking into account the efficacy indicators obtained in the study with a favorable safety profile, the use of BCD-100 in routine clinical practice meets the balance of risk and benefit in this patient population.

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<120> AQUEOUS PHARMACEUTICAL COMPOSITION OF ANTI-PD-1 ANTIBODY PROLGOLIMAB AND ITS APPLICATION

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Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser

165 170 175

Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr

180 185 190

Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser

195 200 205

Phe Asn Arg Gly Glu Cys

210

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Claims (112)

1. An aqueous pharmaceutical composition of an anti-PD-1 antibody, consisting of: (a) prolgolimab at a concentration of 15 mg/ml to 40 mg/ml as an antibody; (b) trehalose dihydrate at a concentration of 80 mg/ml to 110 mg/ml; (c) sodium acetate trihydrate at a concentration of 1.6 mg/ml to 1.9 mg/ml; And (d) acetic acid to pH 5.0-5.5. 2. An aqueous pharmaceutical composition according to claim 1, where said prolgolimab is in a concentration of 15 mg/ml to 25 mg/ml. 3. An aqueous pharmaceutical composition according to claim 2, where said prolgolimab is at a concentration of 20 mg/ml. 4. Aqueous pharmaceutical composition according to any one of paragraphs. 1-3, where the specified trehalose dihydrate is in a concentration of from 95 mg/ml to 105 mg/ml. 5. An aqueous pharmaceutical composition according to claim 4, where said trehalose dihydrate is at a concentration of 100 mg/ml. 6. An aqueous pharmaceutical composition according to claim 1, where said sodium acetate trihydrate is in a concentration of 1.7 mg/ml to 1.8 mg/ml. 7. An aqueous pharmaceutical composition according to claim 6, where said sodium acetate trihydrate is at a concentration of 1.742 mg/ml. 8. Aqueous pharmaceutical composition according to any one of paragraphs. 1-7, where the specified acetic acid is added to pH 5.0. 9. Aqueous pharmaceutical composition of anti-PD-1 antibody, consisting of: (a) prolgolimab at a concentration of 90 mg/ml to 150 mg/ml as an antibody; (b) trehalose dihydrate at a concentration of 75 mg/ml to 85 mg/ml; (c) sodium acetate trihydrate at a concentration of 1.6 mg/ml to 1.9 mg/ml; And (d) acetic acid to a pH of 5.0 to 5.5. 10. An aqueous pharmaceutical composition according to claim 9, where said prolgolimab is in a concentration of 90 mg/ml to 110 mg/ml. 11. An aqueous pharmaceutical composition according to any one of paragraphs. 9, 10, where the specified prolgolimab is at a concentration of 100 mg/ml. 12. An aqueous pharmaceutical composition according to claim 9, where said trehalose dihydrate is at a concentration of 80 mg/ml. 13. An aqueous pharmaceutical composition according to claim 9, where said sodium acetate trihydrate is in a concentration of 1.7 mg/ml to 1.8 mg/ml. 14. An aqueous pharmaceutical composition according to claim 13, where said sodium acetate trihydrate is at a concentration of 1.742 mg/ml. 15. An aqueous pharmaceutical composition of an anti-PD-1 antibody containing: (a) prolgolimab at a concentration of 5 mg/ml to 150 mg/ml as an antibody; (b) trehalose dihydrate at a concentration of 75 mg/ml to 85 mg/ml or at a concentration of 95 mg/ml to 105 mg/ml; (c) L-histidine at a concentration of 0.7 to 1.0 mg/ml; (d) L-histidine hydrochloride at a concentration of 2.8 to 3.3 mg/ml; And the composition has a pH of 5.5 to 6.0. 16. An aqueous pharmaceutical composition according to claim 15, where said prolgolimab is in a concentration of 15 mg/ml to 40 mg/ml. 17. An aqueous pharmaceutical composition according to claim 16, where said prolgolimab is in a concentration of 15 mg/ml to 25 mg/ml. 18. An aqueous pharmaceutical composition according to claim 17, where said prolgolimab is at a concentration of 20 mg/ml. 19. Aqueous pharmaceutical composition according to any one of paragraphs. 16-18, where the specified trehalose dihydrate is in a concentration of from 95 mg/ml to 105 mg/ml. 20. An aqueous pharmaceutical composition according to claim 19, where said trehalose dihydrate is at a concentration of 100 mg/ml. 21. An aqueous pharmaceutical composition according to claim 15, where said prolgolimab is in a concentration of 90 mg/ml to 110 mg/ml. 22. An aqueous pharmaceutical composition according to claim 21, where said prolgolimab is at a concentration of 100 mg/ml. 23. Aqueous pharmaceutical composition according to any one of paragraphs. 21, 22, where the specified trehalose dihydrate is in a concentration of from 75 mg/ml to 85 mg/ml. 24. An aqueous pharmaceutical composition according to claim 23, where said trehalose dihydrate is at a concentration of 80 mg/ml. 25. An aqueous pharmaceutical composition according to claim 15, where said L-histidine is at a concentration of 0.92 mg/ml. 26. An aqueous pharmaceutical composition according to claim 15, where said L-histidine hydrochloride is at a concentration of 2.96 mg/ml. 27. An aqueous pharmaceutical composition according to claim 15, wherein said composition has a pH of 5.5. 28. An aqueous pharmaceutical composition of an anti-PD-1 antibody, consisting of: (a) prolgolimab at a concentration of 20 mg/ml as an antibody; (b) trehalose dihydrate at a concentration of 100 mg/ml; (c) sodium acetate trihydrate at a concentration of 1.7 to 1.8 mg/ml; And (d) acetic acid to pH 5.0. 29. An aqueous pharmaceutical composition of anti-PD-1 antibody according to claim 28, containing: (a) prolgolimab at a concentration of 20 mg/ml as an antibody; (b) trehalose dihydrate at a concentration of 100 mg/ml; (c) sodium acetate trihydrate at a concentration of 1.742 mg/ml; And (d) acetic acid to pH 5.0. 30. An aqueous pharmaceutical composition of an anti-PD-1 antibody, consisting of: (a) prolgolimab at a concentration of 100 mg/ml as an antibody; (b) trehalose dihydrate at a concentration of 80 mg/ml; (c) sodium acetate trihydrate at a concentration of 1.7 mg/ml to 1.8 mg/ml; And (d) acetic acid to a pH of 5.0 to 5.5. 31. An aqueous pharmaceutical composition of anti-PD-1 antibody according to claim 30, containing: (a) prolgolimab at a concentration of 100 mg/ml as an antibody; (b) trehalose dihydrate at a concentration of 80 mg/ml; (c) sodium acetate trihydrate at a concentration of 1.742 mg/ml; And (d) acetic acid to a pH of 5.0 to 5.5. 32. An aqueous pharmaceutical composition of an anti-PD-1 antibody, consisting of: (a) prolgolimab at a concentration of 20 mg/ml as an antibody; (b) trehalose dihydrate at a concentration of 100 mg/ml; (c) L-histidine at a concentration of 0.7 to 1.0 mg/ml; And (d) L-histidine hydrochloride at a concentration of 2.8 to 3.3 mg/ml; (e) wherein said composition has a pH of 5.5. 33. An aqueous pharmaceutical composition of anti-PD-1 antibody according to claim 32, containing: (a) prolgolimab at a concentration of 20 mg/ml as an antibody; (b) trehalose dihydrate at a concentration of 100 mg/ml; (c) L-histidine at a concentration of 0.92 mg/ml; And (d) L-histidine hydrochloride at a concentration of 2.96 mg/ml; (e) wherein said composition has a pH of 5.5. 34. An aqueous pharmaceutical composition of an anti-PD-1 antibody, consisting of: (a) prolgolimab at a concentration of 100 mg/ml as an antibody; (b) trehalose dihydrate at a concentration of 80 mg/ml; (c) L-histidine at a concentration of 0.7 to 1.0 mg/ml; And (d) L-histidine hydrochloride at a concentration of 2.8 to 3.3 mg/ml; (e) wherein said composition has a pH of 5.5 to 6.0. 35. An aqueous pharmaceutical composition of anti-PD-1 antibody according to claim 34, containing: (a) prolgolimab at a concentration of 100 mg/ml as an antibody; (b) trehalose dihydrate at a concentration of 80 mg/ml; (c) L-histidine at a concentration of 0.92 mg/ml; And (d) L-histidine hydrochloride at a concentration of 2.96 mg/ml; (e) wherein said composition has a pH of 5.5 to 6.0. 36. An aqueous pharmaceutical composition of an anti-PD-1 antibody, consisting of: (a) prolgolimab at a concentration of 20 mg as an antibody; (b) trehalose dihydrate at a concentration of 100 mg; (c) sodium acetate trihydrate at a concentration of 1.742 mg; (d) acetic acid to pH 5.0, and (e) water for injection up to 1 ml. 37. Use of an aqueous pharmaceutical composition of the anti-PD-1 antibody prolgolimab according to any one of claims. 1-36 for the treatment of malignancy in a subject in need thereof. 38. Use according to claim 37, where the malignant neoplasm is selected from the group containing melanoma, including inoperable or metastatic melanoma, early stages of melanoma before and after radical treatment; lung cancer, non-small cell lung cancer (NSCLC), including unresectable or metastatic non-small cell lung cancer; non-squamous non-small cell lung cancer, squamous cell lung cancer; small cell lung cancer, including unresectable or metastatic small cell lung cancer; early stage lung cancer before and after radical treatment; cervical cancer, including metastatic cervical cancer, early stages of cervical cancer before and after radical treatment; head and neck tumors, including squamous cell carcinoma of the head and neck; Hodgkin's lymphoma; tumors of the stomach and intestines, metastatic squamous cell carcinoma of the esophagus; bladder cancer, including metastatic urothelial carcinoma, kidney cancer; endometrial cancer, including metastatic endometrial cancer, early stages of endometrial cancer before and after radical treatment; breast cancer, including metastatic breast cancer, early stages of endometrial cancer before and after radical treatment; liver cancer, including metastatic or inoperable liver cancer, early stages of liver cancer before and after radical treatment; unresectable or metastatic solid tumor, including unresectable or metastatic solid tumor with signs of microsatellite instability. 39. Use according to claim 37, where said aqueous pharmaceutical composition is administered at a dose of the anti-PD-1 antibody prolgolimab of 1 mg/kg body weight. 40. Use according to claim 37, where said aqueous pharmaceutical composition is administered at a dose of the anti-PD-1 antibody prolgolimab of 3 mg/kg body weight. 41. Use according to claim 37, wherein said aqueous pharmaceutical composition is administered every 2 weeks. 42. Use according to claim 37, wherein said aqueous pharmaceutical composition is administered every 3 weeks. 43. Use according to claim 37, where the specified aqueous pharmaceutical composition is administered at a dose of the anti-PD-1 antibody prolgolimab 1 mg/kg body weight every 2 weeks. 44. Use according to claim 37, where the specified aqueous pharmaceutical composition is administered at a dose of the anti-PD-1 antibody prolgolimab 3 mg/kg body weight every 3 weeks. 45. Use according to claim 37, wherein said aqueous pharmaceutical composition is administered parenterally. 46. Use according to claim 37, wherein said parenteral administration is intravenous, subcutaneous or intramuscular administration. 47. Use according to claim 37, wherein said aqueous pharmaceutical composition is administered intravenously as an infusion. 48. A method of treating a malignant neoplasm in a subject in need thereof, comprising administering a therapeutically effective amount of an aqueous pharmaceutical composition according to any one of claims. 1-8, 16-19, 28, 29, 32, 33. 49. The method according to claim 48, where the specified aqueous pharmaceutical composition is administered at a dose of the anti-PD-1 antibody prolgolimab 1 mg/kg body weight. 50. The method according to claim 48, where the specified aqueous pharmaceutical composition is administered at a dose of the anti-PD-1 antibody prolgolimab of 3 mg/kg body weight. 51. The method according to claim 48, where the specified aqueous pharmaceutical composition is administered every 2 weeks. 52. The method according to claim 48, where the specified aqueous pharmaceutical composition is administered every 3 weeks. 53. The method according to claim 48, where the specified aqueous pharmaceutical composition is administered at a dose of the anti-PD-1 antibody prolgolimab 1 mg/kg body weight every 2 weeks. 54. The method according to claim 48, where the specified aqueous pharmaceutical composition is administered at a dose of the anti-PD-1 antibody prolgolimab 3 mg/kg body weight every 3 weeks. 55. The method according to claim 48, where the specified aqueous pharmaceutical composition is administered parenterally. 56. The method of claim 55, wherein said parenteral administration is intravenous, subcutaneous or intramuscular. 57. The method according to claim 56, where the specified aqueous pharmaceutical composition is administered intravenously as an infusion. 58. The method according to claim 48, where the malignant neoplasm is melanoma, including inoperable or metastatic melanoma, early stages of melanoma before and after radical treatment; lung cancer, non-small cell lung cancer (NSCLC), including unresectable or metastatic non-small cell lung cancer.

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